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Contents:

PCR primers for off-targets of CCGTCTTCCCCTCCATCGTG GGG

In the table below, Illumina Nextera Handle sequences have been added and highlighted in bold. Primers for the on-target have been added for convenience. The table below is sorted by the CFD off-target score. Sites with very low CFD scores < 0.02 are unlikely to be cleaved, see our study Haeussler et al. 2016, Figure 2.

In the protocol by Matthew Canver, Harvard, two PCRs are run: one PCR to amplify the potential off-target, then a second PCR to extend the handles with Illumina barcodes. Please click here to download the protocol. Alternatively, you can have a look at Fu et al, 2014.

NamePrimer Sequence Tm CFD Score
ontarget_mm0__chr7 5.53 Mbp_chr7_5529549_F TCGTCGGCAGCGTCCTCCCCTACCTGGTGCCT 59.9 1.00
ontarget_mm0__chr7 5.53 Mbp_chr7_5529549_R GTCTCGTGGGCTCGGTGGATGATGATATCGCCGCG 60.4 1.00
mm0__chr11 1.80 Mbp_chr11_1803987_F TCGTCGGCAGCGTCCGCCACCCACATAGAAGTCC 60.4 1.00
mm0__chr11 1.80 Mbp_chr11_1803987_R GTCTCGTGGGCTCGGAACACCTCTGGCATGTGCAA 60.4 1.00
mm1__chr3 175.98 Mbp_chr3_175978471_F TCGTCGGCAGCGTCGAGTCCTTCTGACCCATGCC 60.1 0.75
mm1__chr3 175.98 Mbp_chr3_175978471_R GTCTCGTGGGCTCGGGCAAAGGCTCCTGTATGTGC 59.5 0.75
mm1__chr19 9.52 Mbp_chr19_9518234_F TCGTCGGCAGCGTCAGTTAACAATGGCTCCGGCA 59.9 0.75
mm1__chr19 9.52 Mbp_chr19_9518234_R GTCTCGTGGGCTCGGGGAGTCCTTCTGACCGATGC 60.1 0.75
mm1__chr9 6.83 Mbp_chr9_6834539_F Primer3: not found at this Tm N.d. 0.70
mm1__chr9 6.83 Mbp_chr9_6834539_R Primer3: not found at this Tm N.d. 0.70
mm2__chr18 62.44 Mbp_chr18_62442093_F TCGTCGGCAGCGTCCCTCGTCATCGACAATGGCT 60.1 0.65
mm2__chr18 62.44 Mbp_chr18_62442093_R GTCTCGTGGGCTCGGGAGTCCTTCTGACCCATGCC 60.1 0.65
mm3__chrX 46.29 Mbp_chrX_46289021_F Primer3: not found at this Tm N.d. 0.62
mm3__chrX 46.29 Mbp_chrX_46289021_R Primer3: not found at this Tm N.d. 0.62
mm2__chr5 77.78 Mbp_chr5_77784971_F TCGTCGGCAGCGTCCTCGTCATTGACAACGGCTC 59.2 0.50
mm2__chr5 77.78 Mbp_chr5_77784971_R GTCTCGTGGGCTCGGAATCCTTCTGACCCATGCCC 59.7 0.50
mm1__chr6 88.28 Mbp_chr6_88276899_F TCGTCGGCAGCGTCGAGTCCTTCTGACCCATGCC 60.1 0.46
mm1__chr6 88.28 Mbp_chr6_88276899_R GTCTCGTGGGCTCGGTGGATGATGCTATCACCGCG 60.3 0.46
mm2__chr10 69.02 Mbp_chr10_69022838_F Primer3: not found at this Tm N.d. 0.43
mm2__chr10 69.02 Mbp_chr10_69022838_R Primer3: not found at this Tm N.d. 0.43
mm3__chr1 66.76 Mbp_chr1_66756775_F TCGTCGGCAGCGTCACAAAGTCCACAGACCCCTA 57.8 0.40
mm3__chr1 66.76 Mbp_chr1_66756775_R GTCTCGTGGGCTCGGGAAGCAACATGGGGCAATGG 60.1 0.40
mm4__chr12 126.37 Mbp_chr12_126368846_F Primer3: not found at this Tm N.d. 0.33
mm4__chr12 126.37 Mbp_chr12_126368846_R Primer3: not found at this Tm N.d. 0.33
mm2__chr5 131.66 Mbp_chr5_131660008_F TCGTCGGCAGCGTCATGCCCATCATCACACCCTG 60.1 0.32
mm2__chr5 131.66 Mbp_chr5_131660008_R GTCTCGTGGGCTCGGTGTGGAAGGTCTGCTTCACG 60.2 0.32
mm4__chr3 149.42 Mbp_chr3_149421159_F TCGTCGGCAGCGTCTCTTTTCTCAACAGTGATCCTACCT 59.4 0.31
mm4__chr3 149.42 Mbp_chr3_149421159_R GTCTCGTGGGCTCGGTTGTGACCTCCTCTCCTCCC 60.2 0.31
mm4__chr17 78.17 Mbp_chr17_78171381_F TCGTCGGCAGCGTCGGGACTTTGGAGGTGGCTC 60.0 0.30
mm4__chr17 78.17 Mbp_chr17_78171381_R GTCTCGTGGGCTCGGGTTCCATGGAAAGGGAGGGG 60.0 0.30
mm4__chr20 49.81 Mbp_chr20_49812915_F Primer3: not found at this Tm N.d. 0.29
mm4__chr20 49.81 Mbp_chr20_49812915_R Primer3: not found at this Tm N.d. 0.29
mm2__chr22 15.72 Mbp_chr22_15721571_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr22 15.72 Mbp_chr22_15721571_R GTCTCGTGGGCTCGGATCATGTTCTGGTGCCTGGG 60.0 0.28
mm2__chr2 131.26 Mbp_chr2_131263640_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr2 131.26 Mbp_chr2_131263640_R GTCTCGTGGGCTCGGCTCTTTCTGATGCATGCCCC 58.9 0.28
mm2__chr2 130.66 Mbp_chr2_130656834_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr2 130.66 Mbp_chr2_130656834_R GTCTCGTGGGCTCGGCTCTTTCTGATGCATGCCCC 58.9 0.28
mm2__chr2 130.46 Mbp_chr2_130463835_F TCGTCGGCAGCGTCCTCTTTCTGATGCATGCCCC 58.9 0.28
mm2__chr2 130.46 Mbp_chr2_130463835_R GTCTCGTGGGCTCGGGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr2 130.08 Mbp_chr2_130075263_F TCGTCGGCAGCGTCCTCTTTCTGATGCATGCCCC 58.9 0.28
mm2__chr2 130.08 Mbp_chr2_130075263_R GTCTCGTGGGCTCGGGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr14 19.40 Mbp_chr14_19402800_F TCGTCGGCAGCGTCATCATGTTCTGGTGCCTGGG 60.0 0.28
mm2__chr14 19.40 Mbp_chr14_19402800_R GTCTCGTGGGCTCGGGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr14 19.00 Mbp_chr14_18998952_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.28
mm2__chr14 19.00 Mbp_chr14_18998952_R GTCTCGTGGGCTCGGATCATGTTCTGGTGCCTGGG 60.0 0.28
mm4__chr5 62.19 Mbp_chr5_62190235_F Primer3: not found at this Tm N.d. 0.27
mm4__chr5 62.19 Mbp_chr5_62190235_R Primer3: not found at this Tm N.d. 0.27
mm4__chr7 63.40 Mbp_chr7_63404893_F TCGTCGGCAGCGTCAGGAAACTCATCAGCTTGGGT 59.2 0.26
mm4__chr7 63.40 Mbp_chr7_63404893_R GTCTCGTGGGCTCGGTTCCAGCAAAAGGAGTGGCT 59.8 0.26
mm4__chr7 63.34 Mbp_chr7_63343194_F TCGTCGGCAGCGTCTTCCAGCAAAAGGAGTGGCT 59.8 0.26
mm4__chr7 63.34 Mbp_chr7_63343194_R GTCTCGTGGGCTCGGAGGAAACTCATCAGCTTGGGT 59.2 0.26
mm4__chr11 111.28 Mbp_chr11_111284162_F TCGTCGGCAGCGTCGGTCACGTCAGGTTACTACGG 60.1 0.26
mm4__chr11 111.28 Mbp_chr11_111284162_R GTCTCGTGGGCTCGGGAGGACTCACAGGGAAACGG 60.0 0.26
mm4__chrX 150.20 Mbp_chrX_150201605_F TCGTCGGCAGCGTCAAGGGGCAGTGCAAGATGAT 59.6 0.25
mm4__chrX 150.20 Mbp_chrX_150201605_R GTCTCGTGGGCTCGGCCCTTCCTCATGTGCACACA 60.2 0.25
mm4__chr1 46.44 Mbp_chr1_46442706_F Primer3: not found at this Tm N.d. 0.25
mm4__chr1 46.44 Mbp_chr1_46442706_R Primer3: not found at this Tm N.d. 0.25
mm4__chr18 80.25 Mbp_chr18_80246940_F TCGTCGGCAGCGTCGAATCCGAGCAGCCCCTG 60.2 0.25
mm4__chr18 80.25 Mbp_chr18_80246940_R GTCTCGTGGGCTCGGTTGTTCCACGAGTGAGAGGC 59.9 0.25
mm4__chr20 58.79 Mbp_chr20_58793065_F TCGTCGGCAGCGTCCGTCTTCTCCCGCATCATCA 59.8 0.24
mm4__chr20 58.79 Mbp_chr20_58793065_R GTCTCGTGGGCTCGGAATAACCAGGACAGACGGGG 59.0 0.24
mm4__chr20 58.79 Mbp_chr20_58793033_F TCGTCGGCAGCGTCCGTCTTCTCCCGCATCATCA 59.8 0.24
mm4__chr20 58.79 Mbp_chr20_58793033_R GTCTCGTGGGCTCGGAATAACCAGGACAGACGGGG 59.0 0.24
mm4__chr20 58.79 Mbp_chr20_58792969_F TCGTCGGCAGCGTCGCATCATCAGGCTGCAAGC 59.9 0.24
mm4__chr20 58.79 Mbp_chr20_58792969_R GTCTCGTGGGCTCGGTGATGATGCGGGAGAAGACG 59.8 0.24
mm4__chr20 58.79 Mbp_chr20_58792937_F TCGTCGGCAGCGTCGCATCATCAGGCTGCAAGC 59.9 0.24
mm4__chr20 58.79 Mbp_chr20_58792937_R GTCTCGTGGGCTCGGTGATGATGCGGGAGAAGACG 59.8 0.24
mm4__chr20 58.79 Mbp_chr20_58792905_F TCGTCGGCAGCGTCGCATCATCAGGCTGCAAGC 59.9 0.24
mm4__chr20 58.79 Mbp_chr20_58792905_R GTCTCGTGGGCTCGGTGATGATGCGGGAGAAGACG 59.8 0.24
mm3__chr15 43.99 Mbp_chr15_43990074_F TCGTCGGCAGCGTCGAGTCCTTCTGACCCATGCC 60.1 0.24
mm3__chr15 43.99 Mbp_chr15_43990074_R GTCTCGTGGGCTCGGTGTCGTCGACAATGGCTCTG 60.3 0.24
mm2__chr22 41.08 Mbp_chr22_41075131_F TCGTCGGCAGCGTCGGAGTCCTTCTGACCCATGC 60.1 0.23
mm2__chr22 41.08 Mbp_chr22_41075131_R GTCTCGTGGGCTCGGTGTTGACAACAGCTCTGGCA 60.1 0.23
mm4__chrX 55.66 Mbp_chrX_55658884_F TCGTCGGCAGCGTCAGACCAAGGAACCACACGTG 60.1 0.22
mm4__chrX 55.66 Mbp_chrX_55658884_R GTCTCGTGGGCTCGGGGGTCTGTTCCCCATGTTTCT 59.9 0.22
mm4__chr5 6.18 Mbp_chr5_6180901_F TCGTCGGCAGCGTCTGTGAGCCTCCTCTCCTTCT 59.5 0.22
mm4__chr5 6.18 Mbp_chr5_6180901_R GTCTCGTGGGCTCGGAGTAGAGGAGGGCAACACCT 59.8 0.22
mm4__chr10 126.06 Mbp_chr10_126062426_F TCGTCGGCAGCGTCGAGAGTGGGTGGGAGAGAGT 59.9 0.22
mm4__chr10 126.06 Mbp_chr10_126062426_R GTCTCGTGGGCTCGGCCTGCTTGGGCTCTCATCAT 59.8 0.22
mm3__chr5 6.18 Mbp_chr5_6181269_F Primer3: not found at this Tm N.d. 0.21
mm3__chr5 6.18 Mbp_chr5_6181269_R Primer3: not found at this Tm N.d. 0.21
mm3__chr2 131.63 Mbp_chr2_131626223_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.20
mm3__chr2 131.63 Mbp_chr2_131626223_R GTCTCGTGGGCTCGGACTCCTTCTGATGCATGCCC 60.1 0.20
mm4__chr15 99.75 Mbp_chr15_99750717_F Primer3: not found at this Tm N.d. 0.20
mm4__chr15 99.75 Mbp_chr15_99750717_R Primer3: not found at this Tm N.d. 0.20
mm4__chr15 99.75 Mbp_chr15_99750670_F Primer3: not found at this Tm N.d. 0.20
mm4__chr15 99.75 Mbp_chr15_99750670_R Primer3: not found at this Tm N.d. 0.20
mm4__chr15 99.75 Mbp_chr15_99750623_F Primer3: not found at this Tm N.d. 0.20
mm4__chr15 99.75 Mbp_chr15_99750623_R Primer3: not found at this Tm N.d. 0.20
mm3__chr3 12.07 Mbp_chr3_12070309_F TCGTCGGCAGCGTCTCATTGACAGTGTCTCCGGC 60.0 0.20
mm3__chr3 12.07 Mbp_chr3_12070309_R GTCTCGTGGGCTCGGGAGTCCTTCTGACCCATGCC 60.1 0.20
mm4__chr16 87.38 Mbp_chr16_87383938_F TCGTCGGCAGCGTCTACGTAGAGCTCCGCCCC 60.5 0.18
mm4__chr16 87.38 Mbp_chr16_87383938_R GTCTCGTGGGCTCGGCTTTTCTCATGCCCGTCACG 59.5 0.18
mm4__chr14 84.99 Mbp_chr14_84991950_F TCGTCGGCAGCGTCGGACGAGTTGCTGAATTGTTCA 59.4 0.18
mm4__chr14 84.99 Mbp_chr14_84991950_R GTCTCGTGGGCTCGGGGTCATTAAACGTATCTTTCCGTCA 59.4 0.18
mm4__chr3 50.40 Mbp_chr3_50400014_F TCGTCGGCAGCGTCGGACAAGTACCAGGATGCCC 60.1 0.17
mm4__chr3 50.40 Mbp_chr3_50400014_R GTCTCGTGGGCTCGGTGCACAATACACAGCTAATGCA 58.9 0.17
mm4__chr5 15.45 Mbp_chr5_15450772_F Primer3: not found at this Tm N.d. 0.17
mm4__chr5 15.45 Mbp_chr5_15450772_R Primer3: not found at this Tm N.d. 0.17
mm4__chr2 215.46 Mbp_chr2_215456200_F Primer3: not found at this Tm N.d. 0.17
mm4__chr2 215.46 Mbp_chr2_215456200_R Primer3: not found at this Tm N.d. 0.17
mm3__chr5 6.18 Mbp_chr5_6180838_F TCGTCGGCAGCGTCGTGACCCTCCTCTCCCTCTC 60.3 0.16
mm3__chr5 6.18 Mbp_chr5_6180838_R GTCTCGTGGGCTCGGGAAAGAGGAGGGAGCACCAC 60.0 0.16
mm3__chr5 6.18 Mbp_chr5_6180769_F TCGTCGGCAGCGTCGTGACCCTCCTCTCCCTCTC 60.3 0.16
mm3__chr5 6.18 Mbp_chr5_6180769_R GTCTCGTGGGCTCGGAGGCTCACACCTCCCTCAG 60.6 0.16
mm3__chr5 6.18 Mbp_chr5_6180592_F TCGTCGGCAGCGTCAGGTGTGACCCTCCCTTCTT 60.1 0.16
mm3__chr5 6.18 Mbp_chr5_6180592_R GTCTCGTGGGCTCGGCATCAGGATGGAGGGGAGGA 60.1 0.16
mm4__chrX 53.14 Mbp_chrX_53143804_F TCGTCGGCAGCGTCCATGGCCCATGTAGGAGTCC 59.8 0.16
mm4__chrX 53.14 Mbp_chrX_53143804_R GTCTCGTGGGCTCGGCTCTGGCATGTGCAAAGCTG 60.1 0.16
mm4__chr5 6.18 Mbp_chr5_6180532_F TCGTCGGCAGCGTCCACCCCAGGACTGTCTTTCC 59.9 0.16
mm4__chr5 6.18 Mbp_chr5_6180532_R GTCTCGTGGGCTCGGGAGGCAAAGAAGGGAGGGTC 60.0 0.16
mm4__chr4 5.82 Mbp_chr4_5816710_F Primer3: not found at this Tm N.d. 0.16
mm4__chr4 5.82 Mbp_chr4_5816710_R Primer3: not found at this Tm N.d. 0.16
mm4__chr6 101.43 Mbp_chr6_101431429_F TCGTCGGCAGCGTCTGATGCTGTCCTGTGTGCTT 59.8 0.14
mm4__chr6 101.43 Mbp_chr6_101431429_R GTCTCGTGGGCTCGGCGGCTTTGCTGGGGATGA 60.0 0.14
mm4__chr6 34.55 Mbp_chr6_34554131_F TCGTCGGCAGCGTCGGCGGGACCTCACCTGTT 61.6 0.14
mm4__chr6 34.55 Mbp_chr6_34554131_R GTCTCGTGGGCTCGGTGACTTGTTGCAGCCTGAGT 59.8 0.14
mm4__chr5 6.18 Mbp_chr5_6181146_F TCGTCGGCAGCGTCCTCCACACCTCACCACCATC 60.0 0.14
mm4__chr5 6.18 Mbp_chr5_6181146_R GTCTCGTGGGCTCGGAGCACCACAGGAGAGAAGGA 60.1 0.14
mm4__chr2 6.76 Mbp_chr2_6764457_F Primer3: not found at this Tm N.d. 0.14
mm4__chr2 6.76 Mbp_chr2_6764457_R Primer3: not found at this Tm N.d. 0.14
mm4__chr10 86.34 Mbp_chr10_86341339_F TCGTCGGCAGCGTCTGCACTGCCCACTAAACAGT 59.8 0.13
mm4__chr10 86.34 Mbp_chr10_86341339_R GTCTCGTGGGCTCGGTAAGTACCCAGGGTCCCACC 60.2 0.13
mm3__chr11 15.08 Mbp_chr11_15076495_F TCGTCGGCAGCGTCTGTCTTCATGCTGGTATTGGT 57.2 0.13
mm3__chr11 15.08 Mbp_chr11_15076495_R GTCTCGTGGGCTCGGACAGTCTCAAGAAGCTTCCCT 58.6 0.13
mm4__chr1 57.88 Mbp_chr1_57883441_F TCGTCGGCAGCGTCTGCAAAGCCAGACTGCATTG 59.6 0.13
mm4__chr1 57.88 Mbp_chr1_57883441_R GTCTCGTGGGCTCGGAATTTTGCACAGCACCAGGC 60.2 0.13
mm4__chr5 147.47 Mbp_chr5_147466726_F TCGTCGGCAGCGTCAAGCCAACCCCAAAGTGTGA 60.0 0.13
mm4__chr5 147.47 Mbp_chr5_147466726_R GTCTCGTGGGCTCGGACAACACAGCCAAAGTCCCT 59.7 0.13
mm4__chr12 124.61 Mbp_chr12_124610040_F TCGTCGGCAGCGTCCAGAGGGATAGGACTGGGGT 59.7 0.13
mm4__chr12 124.61 Mbp_chr12_124610040_R GTCTCGTGGGCTCGGTATGTTGTCAGGGTCTGGGC 59.3 0.13
mm4__chr22 45.67 Mbp_chr22_45674072_F Primer3: not found at this Tm N.d. 0.12
mm4__chr22 45.67 Mbp_chr22_45674072_R Primer3: not found at this Tm N.d. 0.12
mm4__chr17 186.79 Kbp_chr17_186793_F Primer3: not found at this Tm N.d. 0.12
mm4__chr17 186.79 Kbp_chr17_186793_R Primer3: not found at this Tm N.d. 0.12
mm3__chr15 79.51 Mbp_chr15_79513650_F Primer3: not found at this Tm N.d. 0.12
mm3__chr15 79.51 Mbp_chr15_79513650_R Primer3: not found at this Tm N.d. 0.12
mm4__chr6 46.21 Mbp_chr6_46205060_F TCGTCGGCAGCGTCCGGCATGTGAAAAGCAGCTT 60.0 0.12
mm4__chr6 46.21 Mbp_chr6_46205060_R GTCTCGTGGGCTCGGCACGTAGGAGTCCTTCTGGC 59.8 0.12
mm4__chr20 50.70 Mbp_chr20_50703600_F Primer3: not found at this Tm N.d. 0.12
mm4__chr20 50.70 Mbp_chr20_50703600_R Primer3: not found at this Tm N.d. 0.12
mm4__chr16 5.91 Mbp_chr16_5905903_F TCGTCGGCAGCGTCCACTCCCCAAATGCCTCCTC 60.3 0.12
mm4__chr16 5.91 Mbp_chr16_5905903_R GTCTCGTGGGCTCGGCCTAAGGTAGGGCTTTGGGC 60.1 0.12
mm4__chr22 29.27 Mbp_chr22_29268594_F Primer3: not found at this Tm N.d. 0.12
mm4__chr22 29.27 Mbp_chr22_29268594_R Primer3: not found at this Tm N.d. 0.12
mm4__chr1 224.74 Mbp_chr1_224737651_F TCGTCGGCAGCGTCGACTCGTGTATGCAGAGCCA 59.8 0.11
mm4__chr1 224.74 Mbp_chr1_224737651_R GTCTCGTGGGCTCGGCCTCAGGGAGCTCAAACTATAAACT 60.1 0.11
mm4__chr3 72.56 Mbp_chr3_72557831_F Primer3: not found at this Tm N.d. 0.10
mm4__chr3 72.56 Mbp_chr3_72557831_R Primer3: not found at this Tm N.d. 0.10
mm4__chr5 6.18 Mbp_chr5_6181207_F TCGTCGGCAGCGTCTCCTTCTCTCCTGTGGTGCT 60.1 0.10
mm4__chr5 6.18 Mbp_chr5_6181207_R GTCTCGTGGGCTCGGGGAGAGAGGAAAGGGCACAC 60.0 0.10
mm4__chr5 133.14 Mbp_chr5_133144675_F Primer3: not found at this Tm N.d. 0.10
mm4__chr5 133.14 Mbp_chr5_133144675_R Primer3: not found at this Tm N.d. 0.10
mm4__chr10 129.19 Mbp_chr10_129192607_F TCGTCGGCAGCGTCGGCTGGCCATTTCCTTCTTG 59.4 0.09
mm4__chr10 129.19 Mbp_chr10_129192607_R GTCTCGTGGGCTCGGACTGTTGCAATGTCCTAGCGA 59.9 0.09
mm4__chr20 57.17 Mbp_chr20_57166973_F TCGTCGGCAGCGTCGGTGACCACTGCTAACCCTC 60.0 0.09
mm4__chr20 57.17 Mbp_chr20_57166973_R GTCTCGTGGGCTCGGTGCACACGGGTAGTCCTTTC 59.9 0.09
mm4__chr11 36.31 Mbp_chr11_36305686_F Primer3: not found at this Tm N.d. 0.09
mm4__chr11 36.31 Mbp_chr11_36305686_R Primer3: not found at this Tm N.d. 0.09
mm3__chr2 27.37 Mbp_chr2_27373995_F TCGTCGGCAGCGTCAGGATGTCCTGGAGGTGAGG 60.3 0.09
mm3__chr2 27.37 Mbp_chr2_27373995_R GTCTCGTGGGCTCGGCAGCTACAGCCTGGGGATAG 59.3 0.09
mm4__chr16 50.06 Mbp_chr16_50059123_F TCGTCGGCAGCGTCCGCCCACATAGGAGTCCTTC 59.8 0.09
mm4__chr16 50.06 Mbp_chr16_50059123_R GTCTCGTGGGCTCGGTGGCTTTGCTAGGGACGATG 60.1 0.09
mm4__chr12 57.97 Mbp_chr12_57966032_F Primer3: not found at this Tm N.d. 0.08
mm4__chr12 57.97 Mbp_chr12_57966032_R Primer3: not found at this Tm N.d. 0.08
mm4__chr16 84.42 Mbp_chr16_84419399_F Primer3: not found at this Tm N.d. 0.08
mm4__chr16 84.42 Mbp_chr16_84419399_R Primer3: not found at this Tm N.d. 0.08
mm4__chr9 16.15 Mbp_chr9_16153481_F TCGTCGGCAGCGTCGCTCAACCCTGGGAACTGAG 60.3 0.08
mm4__chr9 16.15 Mbp_chr9_16153481_R GTCTCGTGGGCTCGGCCATTCACTCGGCTGTGCTC 61.3 0.08
mm4__chr5 6.18 Mbp_chr5_6181024_F Primer3: not found at this Tm N.d. 0.07
mm4__chr5 6.18 Mbp_chr5_6181024_R Primer3: not found at this Tm N.d. 0.07
mm4__chr1 164.85 Mbp_chr1_164852508_F TCGTCGGCAGCGTCTCCCCAGTGTTCTAGCTGGA 59.8 0.07
mm4__chr1 164.85 Mbp_chr1_164852508_R GTCTCGTGGGCTCGGGCATGTGATGGCCCTAGAGT 59.5 0.07
mm4__chr20 61.84 Mbp_chr20_61835977_F TCGTCGGCAGCGTCTGCCAGAAGGACCCTGACTA 59.8 0.07
mm4__chr20 61.84 Mbp_chr20_61835977_R GTCTCGTGGGCTCGGTGGAAACACAGATGGAGCCC 59.9 0.07
mm4__chr12 56.24 Mbp_chr12_56241970_F TCGTCGGCAGCGTCCCATCTTTCCTCCCTGCTGG 60.1 0.07
mm4__chr12 56.24 Mbp_chr12_56241970_R GTCTCGTGGGCTCGGGACAAGGGACCGTGGCTAC 60.0 0.07
mm4__chr17 81.51 Mbp_chr17_81512246_F TCGTCGGCAGCGTCCCATCCACTCACCTGGTGTC 60.0 0.06
mm4__chr17 81.51 Mbp_chr17_81512246_R GTCTCGTGGGCTCGGATGGAAGAAGAGATCGCCGC 60.2 0.06
mm3__chr10 71.74 Mbp_chr10_71742343_F TCGTCGGCAGCGTCCAGAAAGGGTGCCTCTCTGG 60.0 0.06
mm3__chr10 71.74 Mbp_chr10_71742343_R GTCTCGTGGGCTCGGACCCTGACGTGTTGTGACAG 60.1 0.06
mm4__chr1 110.21 Mbp_chr1_110209706_F TCGTCGGCAGCGTCGCCAGAGCTACGGTTTCCTT 60.0 0.06
mm4__chr1 110.21 Mbp_chr1_110209706_R GTCTCGTGGGCTCGGATGGTCTTGATGCTCTCCGC 60.1 0.06
mm4__chr2 241.32 Mbp_chr2_241315785_F TCGTCGGCAGCGTCCAACTGCCGTGGGAGGAG 60.0 0.06
mm4__chr2 241.32 Mbp_chr2_241315785_R GTCTCGTGGGCTCGGAACTGCTTCCTGGCTCAGTG 60.2 0.06
mm4__chr8 96.32 Mbp_chr8_96322385_F Primer3: not found at this Tm N.d. 0.06
mm4__chr8 96.32 Mbp_chr8_96322385_R Primer3: not found at this Tm N.d. 0.06
mm4__chr2 96.13 Mbp_chr2_96131842_F TCGTCGGCAGCGTCTGAAAGGTCTCGTGCTGGAC 59.9 0.06
mm4__chr2 96.13 Mbp_chr2_96131842_R GTCTCGTGGGCTCGGTGGGCTTGATGGTCTGGTTC 59.9 0.06
mm4__chr15 87.95 Mbp_chr15_87953912_F TCGTCGGCAGCGTCACACATTGTCTGCAGGCTGT 60.1 0.05
mm4__chr15 87.95 Mbp_chr15_87953912_R GTCTCGTGGGCTCGGGCTGGCAATGGAGGTCGATA 59.8 0.05
mm4__chr10 124.53 Mbp_chr10_124526823_F TCGTCGGCAGCGTCCCAGAAGTCCCATCGTTGCT 60.0 0.05
mm4__chr10 124.53 Mbp_chr10_124526823_R GTCTCGTGGGCTCGGGGCTTCCCAGATCTGCCTG 60.1 0.05
mm4__chr19 10.09 Mbp_chr19_10086076_F TCGTCGGCAGCGTCGAGGAGGGTGATGAACTGGG 59.4 0.05
mm4__chr19 10.09 Mbp_chr19_10086076_R GTCTCGTGGGCTCGGTGCAGTCGGTCACATGTCC 60.0 0.05
mm4__chr1 208.02 Mbp_chr1_208016929_F TCGTCGGCAGCGTCAGGCAGAGGAGAAGTCAGGT 59.8 0.05
mm4__chr1 208.02 Mbp_chr1_208016929_R GTCTCGTGGGCTCGGCCCCACAGAAGAGCCTACAG 59.4 0.05
mm4__chr12 76.72 Mbp_chr12_76715361_F Primer3: not found at this Tm N.d. 0.05
mm4__chr12 76.72 Mbp_chr12_76715361_R Primer3: not found at this Tm N.d. 0.05
mm4__chr1 181.48 Mbp_chr1_181481588_F TCGTCGGCAGCGTCGTTTCCTTCCTCAGCCCCTC 60.0 0.05
mm4__chr1 181.48 Mbp_chr1_181481588_R GTCTCGTGGGCTCGGCTGTGTTTTCCTGCCTTGGC 59.9 0.05
mm4__chr10 7.64 Mbp_chr10_7638220_F Primer3: not found at this Tm N.d. 0.05
mm4__chr10 7.64 Mbp_chr10_7638220_R Primer3: not found at this Tm N.d. 0.05
mm4__chr17 3.92 Mbp_chr17_3915975_F TCGTCGGCAGCGTCCCTCGTCCCCATCTCAACAG 59.8 0.04
mm4__chr17 3.92 Mbp_chr17_3915975_R GTCTCGTGGGCTCGGTCTGACATCCCTTGCGTGAC 60.0 0.04
mm4__chr3 143.86 Mbp_chr3_143855297_F TCGTCGGCAGCGTCGGAAAGCCGAGGACAGCATA 59.8 0.04
mm4__chr3 143.86 Mbp_chr3_143855297_R GTCTCGTGGGCTCGGCTGCTGTCTCTGCAAAAGGC 59.7 0.04
mm4__chr1 15.74 Mbp_chr1_15742593_F TCGTCGGCAGCGTCGGGCTGCCATGTTCTCTCTT 60.0 0.04
mm4__chr1 15.74 Mbp_chr1_15742593_R GTCTCGTGGGCTCGGGGAGGGAGTCAAGGAGGCTA 60.0 0.04
mm4__chr22 17.02 Mbp_chr22_17015514_F TCGTCGGCAGCGTCATGCTCAAAAGGAAGGGCCA 59.8 0.04
mm4__chr22 17.02 Mbp_chr22_17015514_R GTCTCGTGGGCTCGGTCCTCTGCAGGACCTTCTCT 59.5 0.04
mm4__chr22 11.02 Mbp_chr22_11017397_F TCGTCGGCAGCGTCTCCTCTGCAGGACCTTCTCT 59.5 0.04
mm4__chr22 11.02 Mbp_chr22_11017397_R GTCTCGTGGGCTCGGGAGTCCCCTGAACGCTCAAA 59.9 0.04
mm4__chr21 10.36 Mbp_chr21_10364477_F TCGTCGGCAGCGTCTCCTCTGCAGGACCTTCTCT 59.5 0.04
mm4__chr21 10.36 Mbp_chr21_10364477_R GTCTCGTGGGCTCGGGAGTCCCCTGAACGCTCAAA 59.9 0.04
mm4__chr8 6.79 Mbp_chr8_6789194_F Primer3: not found at this Tm N.d. 0.04
mm4__chr8 6.79 Mbp_chr8_6789194_R Primer3: not found at this Tm N.d. 0.04
mm4__chr18 59.03 Mbp_chr18_59026393_F Primer3: not found at this Tm N.d. 0.04
mm4__chr18 59.03 Mbp_chr18_59026393_R Primer3: not found at this Tm N.d. 0.04
mm4__chr22 50.44 Mbp_chr22_50438048_F TCGTCGGCAGCGTCCTCGGAACAGTCGCTTTCCT 60.0 0.04
mm4__chr22 50.44 Mbp_chr22_50438048_R GTCTCGTGGGCTCGGAGAGGGGCTTAGTTCGGGAT 60.0 0.04
mm4__chr15 45.19 Mbp_chr15_45186911_F Primer3: not found at this Tm N.d. 0.03
mm4__chr15 45.19 Mbp_chr15_45186911_R Primer3: not found at this Tm N.d. 0.03
mm4__chr10 129.46 Mbp_chr10_129461459_F TCGTCGGCAGCGTCCCTGCCTTTCCTTCTCTGGG 60.0 0.03
mm4__chr10 129.46 Mbp_chr10_129461459_R GTCTCGTGGGCTCGGTTCTTGGTGAGGGCAGTGTC 59.8 0.03
mm4__chrX 72.02 Mbp_chrX_72017129_F TCGTCGGCAGCGTCAAACAACCCACCCCAGAAGG 60.1 0.03
mm4__chrX 72.02 Mbp_chrX_72017129_R GTCTCGTGGGCTCGGCAATAGGCCCTGCTACCCAG 59.8 0.03
mm4__chr19 12.99 Mbp_chr19_12985900_F TCGTCGGCAGCGTCGAGATCTGATGCCAGCTGGG 60.2 0.03
mm4__chr19 12.99 Mbp_chr19_12985900_R GTCTCGTGGGCTCGGGCTGCATCTAGAGGCCACTC 60.2 0.03
mm4__chr14 69.15 Mbp_chr14_69151683_F Primer3: not found at this Tm N.d. 0.03
mm4__chr14 69.15 Mbp_chr14_69151683_R Primer3: not found at this Tm N.d. 0.03
mm4__chr11 117.48 Mbp_chr11_117483287_F TCGTCGGCAGCGTCATGAACACACATGGGGCACT 59.8 0.03
mm4__chr11 117.48 Mbp_chr11_117483287_R GTCTCGTGGGCTCGGTCCTCCAGTTCATGTTGCCG 60.3 0.03
mm4__chr1 181.38 Mbp_chr1_181375729_F Primer3: not found at this Tm N.d. 0.03
mm4__chr1 181.38 Mbp_chr1_181375729_R Primer3: not found at this Tm N.d. 0.03
mm4__chr21 36.49 Mbp_chr21_36493842_F TCGTCGGCAGCGTCGAACACATGTGGCTGGAGGA 59.9 0.02
mm4__chr21 36.49 Mbp_chr21_36493842_R GTCTCGTGGGCTCGGTGCCCAGCCACTCTATACCT 60.0 0.02
mm4__chr5 61.39 Mbp_chr5_61390720_F TCGTCGGCAGCGTCTTTAAGACCCCACCAGCTGC 60.2 0.02
mm4__chr5 61.39 Mbp_chr5_61390720_R GTCTCGTGGGCTCGGAAGTTTGGCCATGGCTGCTT 61.1 0.02
mm4__chr8 70.40 Mbp_chr8_70402448_F TCGTCGGCAGCGTCGCTCCAGTCCCCGATTCC 59.4 0.02
mm4__chr8 70.40 Mbp_chr8_70402448_R GTCTCGTGGGCTCGGGTCGGTCCCTGGTTAGATGG 59.5 0.02
mm4__chr19 14.51 Mbp_chr19_14506115_F TCGTCGGCAGCGTCAACAAGCTCCTGTCCCTTGG 59.8 0.01
mm4__chr19 14.51 Mbp_chr19_14506115_R GTCTCGTGGGCTCGGGACCGAGCCACTTCCCATAG 59.8 0.01
mm4__chrX 28.51 Mbp_chrX_28505599_F Primer3: not found at this Tm N.d. 0.01
mm4__chrX 28.51 Mbp_chrX_28505599_R Primer3: not found at this Tm N.d. 0.01
mm3__chr1 223.86 Mbp_chr1_223864743_F TCGTCGGCAGCGTCTGACCCATGCTCACCATCAC 60.0 0.01
mm3__chr1 223.86 Mbp_chr1_223864743_R GTCTCGTGGGCTCGGCGCTCGTCATTGACAACAGC 60.1 0.01
mm4__chr9 91.48 Mbp_chr9_91484575_F TCGTCGGCAGCGTCTTTTCCTCTACCCTGCTGCC 59.6 0.01
mm4__chr9 91.48 Mbp_chr9_91484575_R GTCTCGTGGGCTCGGGCACATCCAGGATGCCCATA 59.8 0.01
mm4__chr2 218.28 Mbp_chr2_218283001_F TCGTCGGCAGCGTCTGGCCTTGCTAGGGAAGAGA 60.2 0.01
mm4__chr2 218.28 Mbp_chr2_218283001_R GTCTCGTGGGCTCGGTAGCACTGCACCTGGAAGAC 59.6 0.01
mm4__chr11 67.44 Mbp_chr11_67442136_F TCGTCGGCAGCGTCTCGGGTAGGCCTTGTCAATG 59.7 0.01
mm4__chr11 67.44 Mbp_chr11_67442136_R GTCTCGTGGGCTCGGCATCTGGGCCACCTGTGAG 60.0 0.01
mm4__chr8 31.64 Mbp_chr8_31640555_F Primer3: not found at this Tm N.d. 0.01
mm4__chr8 31.64 Mbp_chr8_31640555_R Primer3: not found at this Tm N.d. 0.01
mm4__chr3 184.16 Mbp_chr3_184156562_F TCGTCGGCAGCGTCGCCAGTGGAGAGACGTTAGG 59.8 0.01
mm4__chr3 184.16 Mbp_chr3_184156562_R GTCTCGTGGGCTCGGTCCAAACTGTGCCTGTCCAG 60.1 0.01
mm4__chr2 89.86 Mbp_chr2_89856316_F Primer3: not found at this Tm N.d. 0.01
mm4__chr2 89.86 Mbp_chr2_89856316_R Primer3: not found at this Tm N.d. 0.01
mm4__chr2 89.33 Mbp_chr2_89326266_F Primer3: not found at this Tm N.d. 0.01
mm4__chr2 89.33 Mbp_chr2_89326266_R Primer3: not found at this Tm N.d. 0.01
mm4__chr1 162.02 Mbp_chr1_162019365_F Primer3: not found at this Tm N.d. 0.00
mm4__chr1 162.02 Mbp_chr1_162019365_R Primer3: not found at this Tm N.d. 0.00
mm4__chr2 236.77 Mbp_chr2_236767634_F Primer3: not found at this Tm N.d. 0.00
mm4__chr2 236.77 Mbp_chr2_236767634_R Primer3: not found at this Tm N.d. 0.00
mm4__chr19 54.14 Mbp_chr19_54141815_F TCGTCGGCAGCGTCTGACTACTCCAGGGTTCCCT 59.2 0.00
mm4__chr19 54.14 Mbp_chr19_54141815_R GTCTCGTGGGCTCGGAGACTTCTGAGATACCAGCGG 58.9 0.00
mm4__chr20 62.59 Mbp_chr20_62590954_F TCGTCGGCAGCGTCATTCTGGTTCCCTGAGCAGC 60.0 0.00
mm4__chr20 62.59 Mbp_chr20_62590954_R GTCTCGTGGGCTCGGGCAATGCACTTGGGGACATG 60.1 0.00

Off-target amplicon sequences

Primers underlined, off-targets in bold.

ontarget_mm0__chr7 5.53 Mbp_chr7_5529549 CTCCCCTACCTGGTGCCTGGGGCGCCCCACGATGGAGGGGAAGACGGCCCGGGGGGCATCGTCGCCCGCGAAGCCGGCCT
TGCACATGCCGGAGCCGTTGTCGACGACGAGCGCGGCGATATCATCATCCA
mm0__chr11 1.80 Mbp_chr11_1803987 CGCCACCCACATAGAAGTCCTTCTGACCCATGCCCACCGTCATGCCCTGGTGTCTGGGGCACCCCACGATGGAGGGGAAG
ACGG
CCCAGGGGGCATCGTCGTCGGTGAAGCTGGCCTTGCACATGCCAGAGGTGTT
mm1__chr3 175.98 Mbp_chr3_175978471 GAGTCCTTCTGACCCATGCCCACCATCACACCTTGGTGCCTGGGGCGCCCCACGATGGATGGGAAGACGGCCCCGGAGGC
ATCGTCGCCTGCGAAGCTGGTCTTGCACATACAGGAGCCTTTGC
mm1__chr19 9.52 Mbp_chr19_9518234 AGTTAACAATGGCTCCGGCATGTGCAAGGTCAACTTCGTGGGCGACAATGCCCCCCGGGCCATCTTCCCCTCCATCGTGG
GG
CATCTGAAGCACCAGGGCGTTATGGTGGGCATCGGTCAGAAGGACTCC
mm2__chr18 62.44 Mbp_chr18_62442093 CCTCGTCATCGACAATGGCTCTGGCATGTGCAGGGCTGGCTTCGCAGGCGACAGTGCTCTCCAGGCCGTCTTCCTCTCCA
TCATGGGG
CACCCCAGGCACCAGGGTGTGATGGTGGGCATGGGTCAGAAGGACTC
mm2__chr5 77.78 Mbp_chr5_77784971 CTCGTCATTGACAACGGCTCCGGCATGTGCAAGGCCAGCTTCACGGGCGACAATGCCGCCCGGGCAGTCTTCCCCTCCAT
CGTTGGG
CACCCCAGGCACCAGGGCGTGATGGTGGGCATGGGTCAGAAGGATT
mm1__chr6 88.28 Mbp_chr6_88276899 GAGTCCTTCTGACCCATGCCCACCATCACTCCCTGGTGCCTAGGGTGCCCCACAATGGAGGGGAAGACGGCCTGGGGAGC
CTTGCGCATGCTGGAGCAGTTGTCGACGACGACGAGCGCGGTGATAGCATCATCCA
mm3__chr1 66.76 Mbp_chr1_66756775 ACAAAGTCCACAGACCCCTATCTACTTCCATAGCACCATCTTCCCCACCATCTTGCGGGCATCATCCCCTCCAGCTATAC
TCAGTTATTCCTCAGTGCTCTATCTGGCCCCCAGCCATTGCCCCATGTTGCTTC
mm2__chr5 131.66 Mbp_chr5_131660008 ATGCCCATCATCACACCCTGGTGCCTCGGGTGCCCCATTATGGAGGGGAAGACGGCCCGGGGGCATCGTTGCCCGTGAAG
CAGACCTTCCACA
mm4__chr3 149.42 Mbp_chr3_149421159 TCTTTTCTCAACAGTGATCCTACCTTATATAATTTTGTATGTGTATTCTTTCACTAAAGTATATCTAAAGCATTTTCCCC
ACGATGGGGAGGAAGTTGG
ATCCACCAGTTGGTGGGGAGGAGAGGAGGTCACAA
mm4__chr17 78.17 Mbp_chr17_78171381 GGGACTTTGGAGGTGGCTCCCGGCCCGGCTTCCAAGTCCTCCCCTCCATAGTGTGGAGGCTCCCCCGGGAGGTCCCTGCC
CTACCTGCCCGCGTCCCCTCCCAGAGTCCTGGAAAGCCCCTCCCTTTCCATGGAAC
mm2__chr22 15.72 Mbp_chr22_15721571 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCAGGTGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGG
TGCCCCAGGCACCAGAACATGAT
mm2__chr2 131.26 Mbp_chr2_131263640 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCGGGCGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGG
CGCCCCAGGCAGCAGGGCATGATGGGGGGCATGCATCAGAAAGAG
mm2__chr2 130.66 Mbp_chr2_130656834 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCGGGCGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGG
TGCCCCAGGCAGCAGGGCATGATGGGGGGCATGCATCAGAAAGAG
mm2__chr2 130.46 Mbp_chr2_130463835 CTCTTTCTGATGCATGCCCCCCATCATGCCCTGCTGCCTGGGGCGCCCCACGATGGAAGGGAAGACAGCCCGGGGGGCAT
CGTCGCCCGCAAAGCCGGCCTTGCACATGCCAGAGCCGTTGTCAATGACGAGCAC
mm2__chr2 130.08 Mbp_chr2_130075263 CTCTTTCTGATGCATGCCCCCCATCATGCCCTGCTGCCTGGGGCGCCCCACGATGGAAGGGAAGACAGCCCGGGGGGCAT
CGTCGCCCGCAAAGCCGGCCTTGCACATGCCAGAGCCGTTGTCAATGACGAGCAC
mm2__chr14 19.40 Mbp_chr14_19402800 ATCATGTTCTGGTGCCTGGGGCACCCCACGATGGAAGGGAAGACAGCCCGGGGGGCATCGTCACCTGCAAAGCCGGCCTT
GCACATGCCAGAGCCGTTGTCAATGACGAGCAC
mm2__chr14 19.00 Mbp_chr14_18998952 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCAGGTGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGG
TGCCCCAGGCACCAGAACATGAT
mm4__chr7 63.40 Mbp_chr7_63404893 AGGAAACTCATCAGCTTGGGTTTGATAAAAGGACGATTTAGAGGTATTCCCCTCCATCTTGGGGAGCGACCCTTGCAGCA
GAAAAATTGTGGTGAGAGTCTGGGGCAAATGGAGCCACTCCTTTTGCTGGAA
mm4__chr7 63.34 Mbp_chr7_63343194 TTCCAGCAAAAGGAGTGGCTCCATTTGCCCCAGACTCTCACCACAATTTTTCTGCTGCAAGGGTCGCTCCCCAAGATGGA
GGGGAATACCT
CTAAATCGTCCTTTTATCAAACCCAAGCTGATGAGTTTCCT
mm4__chr11 111.28 Mbp_chr11_111284162 GGTCACGTCAGGTTACTACGGTGTCAGAAGATCTTTCTTATCTGACTCAGACTTCCACAACAGTAAACAGTTTTCAAATG
ACGTCTACACCTCCAGCGTGGGGAAGCCGTTTCCCTGTGAGTCCTC
mm4__chrX 150.20 Mbp_chrX_150201605 AAGGGGCAGTGCAAGATGATGCTCTGGGCATGCCAGTAGGGGCTCAGCCAAAAGATGGAGGGGAGCACGGAGGCCACTCG
CAGGCAGAAGGAACTGTGTGTGCACATGAGGAAGGG
mm4__chr18 80.25 Mbp_chr18_80246940 GAATCCGAGCAGCCCCTGGCCCTCAACTCGCCCCGGAAAGGCCGCCCTCCCCTCCAATGTGTGGCGCGCCCCGATCGCCT
CTCACTCGTGGAACAA
mm4__chr20 58.79 Mbp_chr20_58793065 CGTCTTCTCCCGCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATC
ATCAGG
CTGCAAGCCCCGTCTGTCCTGGTTATT
mm4__chr20 58.79 Mbp_chr20_58793033 CGTCTTCTCCCGCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATC
ATCAGGCTGCAAGCCCCGTCTGTCCTGGTTATT
mm4__chr20 58.79 Mbp_chr20_58792969 GCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCA
AGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCGCATCATCA
mm4__chr20 58.79 Mbp_chr20_58792937 GCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCA
AGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCGCATCATCA
mm4__chr20 58.79 Mbp_chr20_58792905 GCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCCCATCATCAGGCTGCA
AGCCCCGTCTTCTCCCCCATCATCAGGCTGCAAGCCCCGTCTTCTCCCGCATCATCA
mm3__chr15 43.99 Mbp_chr15_43990074 GAGTCCTTCTGACCCATGCCCACCGTCACGCCCTGGTGTCTGGGGCGCCCCACAATGAAGGGGAAGATGGCCCGGGGTGG
GCATCGTCACCTGCAAAGCTGGCCTTGCACATGCCAGAGCCATTGTCGACGACA
mm2__chr22 41.08 Mbp_chr22_41075131 GGAGTCCTTCTGACCCATGCCCACCATCATGACCTGGTGCCTGGTGCACCCCACGATGGAAGGGAAGATGGCCCAGGGGA
CATCGTCACCCGCGAAGCCGGCCTTGCACATGCCAGAGCTGTTGTCAACA
mm4__chrX 55.66 Mbp_chrX_55658884 AGACCAAGGAACCACACGTGGTATGTTCCATGGAGGCTACCACACTGTGGTGGGGAAGACAGACAGTTAAACTGAAGAGC
AGTTGAACTGCAAAGGAGAAACATGGGGAACAGACCC
mm4__chr5 6.18 Mbp_chr5_6180901 TGTGAGCCTCCTCTCCTTCTCTTCTGTGGTGCTCCCTCCTCTTTCCCTCCATCTTGAGGAAGGTGTTGCCCTCCTCTACT
mm4__chr10 126.06 Mbp_chr10_126062426 GAGAGTGGGTGGGAGAGAGTAGAGCTAAGCCAGGAGCTCCCAGGGAAGCCCCCTCCTTAGGAAGAGGACGTGGCTACCTG
GGTGCTCAGACCTCCAGCCTCCCCCTCCATCATGGGGATGATGAGAGCCCAAGCAGG
mm3__chr2 131.63 Mbp_chr2_131626223 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCGGGCGACGATGCCCCCCAGGCTGTCTTCCCTTC
TATCGTGGGG
CGCCCCAGGCACCAGGGCATGATGGAGGGCATGCATCAGAAGGAGT
mm3__chr3 12.07 Mbp_chr3_12070309 TCATTGACAGTGTCTCCGGCATGTACAAAGCTGGATTTGCTGGGGACGACGCCCCCTGAGCCGTGTTCCCCTCCATCCTT
AGG
CGCCCCTGGCACCAGGGCACGATGGTGGGCATGGGTCAGAAGGACTC
mm4__chr16 87.38 Mbp_chr16_87383938 TACGTAGAGCTCCGCCCCGGCCGCGCCACCCCCTCCCCGCGGGGCGGCGACCTCAGAGACCCCGCACTGCTGCCCTCCCC
TCCAGCGTGAGG
GCGCCGCCCGTGACGGGCATGAGAAAAG
mm4__chr14 84.99 Mbp_chr14_84991950 GGACGAGTTGCTGAATTGTTCAGAAACTGTGTAAATAGAATGTGTTGAAGTTAGTATATAATTTTCTACCATCCGCCCCC
ACGGTGGAGTGGAAGCAGG
TAATAATTTTTTGTGACGGAAAGATACGTTTAATGACC
mm4__chr3 50.40 Mbp_chr3_50400014 GGACAAGTACCAGGATGCCCAGCAGCCCCTGCCTAGCCCAGCACTGGCCCCATTGCCACCCGTCTTCCCTCCCAACTTGT
GG
GTCCTGCATTAGCTGTGTATTGTGCA
mm3__chr5 6.18 Mbp_chr5_6180838 GTGACCCTCCTCTCCCTCTCTTCTGAGGGGNNNNNNNNNNNNNNNNNNNNNNNNNATCCTGAGGGAGGTGTGAGCCTCCT
CTCCTTCTCTTCTGTGGTGCTCCCTCCTCTTTC
mm3__chr5 6.18 Mbp_chr5_6180769 GTGACCCTCCTCTCCCTCTCTCCTATAGTGCTCCCCTCCTCCCCTCCATCCTGAGGGAGGTGTGACCCTCCTCTCCCTCT
CTTCTGAGGGGNNNNNNNNNNNNNNNNNNNNNNNNNATCCTGAGGGAGGTGTGAGCCT
mm3__chr5 6.18 Mbp_chr5_6180592 AGGTGTGACCCTCCCTTCTTTGCCTCCGGTGGTGCCCCCTCCTCCCCTCCATCCTGAGGGAGGTGTGACCCCCTCTCCTT
CTCTCATGCAATGCTTCCCCCTCCTCCCCTCCATCCTGATG
mm4__chrX 53.14 Mbp_chrX_53143804 CATGGCCCATGTAGGAGTCCTTCTGGCCCATGCCCACCATCACACCCTGGTGCCGGGGTCACCTGATGATGGAGGGGAAC
ACAG
CTCAGGGGCCATCCATCCCCAGCAAAGCCAGCTTTGCACATGCCAGAG
mm4__chr5 6.18 Mbp_chr5_6180532 CACCCCAGGACTGTCTTTCCTACACAATGCCTGCTCCTCTTTCCCTCCATCCTGAGGGAGGTGTGACCCTCCCTTCTTTG
CCTC
mm4__chr6 101.43 Mbp_chr6_101431429 TGATGCTGTCCTGTGTGCTTACTAGAAAGACAAGTCTTTACTCTGACAGAGTTTATATTCTGGTGCTGGGGGCGCCCAAC
AATGGAGGGGAACATGG
CTTGGGGGCGTCATCCCCAGCAAAGCCG
mm4__chr6 34.55 Mbp_chr6_34554131 GGCGGGACCTCACCTGTTCCTTCTTGCCCTCCACCTTGAGGGTCTGTGTCCCCCTGTCAGACTAAATTAACTTCCCCGGG
TTATACTCAGGCTGCAACAAGTCA
mm4__chr5 6.18 Mbp_chr5_6181146 CTCCACACCTCACCACCATCCTGAGGGAAATGTGACCCTCCTCTCTTCTCTCCTGTGGTGCTCCCCCTCCTCACCTCCAT
CCTGAGG
AAGGTGTGACCCTCCTCTCCTTCTCTCCTGTGGTGCT
mm4__chr10 86.34 Mbp_chr10_86341339 TGCACTGCCCACTAAACAGTCCCGGAGGATGGAGGAGAAGATGGGACTAGGATGAGGCCCTGGTGGGACCCTGGGTACTT
A
mm3__chr11 15.08 Mbp_chr11_15076495 TGTCTTCATGCTGGTATTGGTATTTTTGCTTATGACAGAGAATGTTTTGAAGACCTCAGGATGGAAGGGAAGACAGCAGG
ACTTACTGAACACATTAGAGATAAAAGAAAATAAGGGAAGCTTCTTGAGACTGT
mm4__chr1 57.88 Mbp_chr1_57883441 TGCAAAGCCAGACTGCATTGTATTCTTAAATTGGGCCTGAAATAGCCACACGATGAAGGGTAATAAGGCAGAGAGCACAA
CCCAGAGTGCCTGGTGCTGTGCAAAATT
mm4__chr5 147.47 Mbp_chr5_147466726 AAGCCAACCCCAAAGTGTGAGGCTGCAATCCCACTTTGTATTTTGCAGGCTTCCCCTCCATCTTCAGGAAGGGACTTTGG
CTGTGTTGT
mm4__chr12 124.61 Mbp_chr12_124610040 CAGAGGGATAGGACTGGGGTCCCGGAGGATGGAGGGGAAGACCCCAGACAGGGGGCCCCAAGCTAAATCTAGCAGAGGGG
CCCAGACCCTGACAACATA
mm4__chr6 46.21 Mbp_chr6_46205060 CGGCATGTGAAAAGCAGCTTTGCTGGGACAATGCCCTCCGAGCCATATTCCCCTCCATCATCGGGCACCCCCGGCACCAG
GGCGTGATGGTGGGCATGGGCCAGAAGGACTCCTACGTG
mm4__chr16 5.91 Mbp_chr16_5905903 CACTCCCCAAATGCCTCCTCCGGAAGCCTTCCTAATTTCCCTCTTTAGAATTCATCTCTTTTTCCCGACGATGGAGGATA
AGAAGG
TTTTCAGCTCTCTTGTAGTGGGAATTGGCTGCCCAAAGCCCTACCTTAGG
mm4__chr1 224.74 Mbp_chr1_224737651 GACTCGTGTATGCAGAGCCATGTCTGAACTGAGCTCAGGGCTAGAGACCCACACAACAGCGGGGAAGACGGGGTTGATAG
TTTATAGTTTGAGCTCCCTGAGG
mm4__chr5 6.18 Mbp_chr5_6181207 TCCTTCTCTCCTGTGGTGCTCCCCTTCTCCCCTCCATCCTGAGGGAGGTGTGACCCTTCTCTCCCTCTCTCCTATGGTGC
TCTCCTTCCTCCCCTCCATCCTGAGGGAGGTGTGCCCTTTCCTCTCTCC
mm4__chr10 129.19 Mbp_chr10_129192607 GGCTGGCCATTTCCTTCTTGTCCATCAGGCCGTCTCCACCTCCATCTGGGGGGCAGATTGGATCTTGCCAGGTTTTAGAT
CTCAAGCCCGACATTTAGTAGCACTCAATAAAGATTCGCTAGGACATTGCAACAGT
mm4__chr20 57.17 Mbp_chr20_57166973 GGTGACCACTGCTAACCCTCCCAGCCCCCCAGCACGGTCCACACACTATTGTCACCAGGCGATGGAGGAGAAGACAGAGG
GAAAGGACTACCCGTGTGCA
mm3__chr2 27.37 Mbp_chr2_27373995 AGGATGTCCTGGAGGTGAGGCGCTTGTTCAGCACTGCCCCTTCTTCCCCTACATCCTGGGGTCCTGGGCTTGGAGAATGA
TTGGAACCAGCCAGTATGATGAGCTGTACTTCTATCCCTATCCCCAGGCTGTAGCTG
mm4__chr16 50.06 Mbp_chr16_50059123 CGCCCACATAGGAGTCCTTCTGGCCCATCCATGCCCACCATCATGAGCTGGTGCCCGGGGCACCCGACAATGGAGGGGAA
CATGG
TTCAGGGGGCATCGTCCCTAGCAAAGCCA
mm4__chr9 16.15 Mbp_chr9_16153481 GCTCAACCCTGGGAACTGAGGTCCGTTTTCCCCTCCTTCTTAAGGACCATCTCCCTCTAATAATTTGCAGAGANNNNNNN
NNNNNNNNNNNNNNNNNNNNGAGCACAGCCGAGTGAATGG
mm4__chr1 164.85 Mbp_chr1_164852508 TCCCCAGTGTTCTAGCTGGAACACCAGGCCTCCACACCTAAGTCTGTGCCCAAGTCTTTAACACCATCTTCCCATCCATA
GAGTGG
GATGGAACTCCTTGTCTGCCTTTTGACATGTTACTCTAGGGCCATCACATGC
mm4__chr20 61.84 Mbp_chr20_61835977 TGCCAGAAGGACCCTGACTACCCACAGGCTGGAGGGGAAGGCCGGCCGGTTGGGTGATCCCAGGGCTCTGGGTCTTGACC
GGGCTCTGGCATTGCAAGGAAGGGCTCCATCTGTGTTTCCA
mm4__chr12 56.24 Mbp_chr12_56241970 CCATCTTTCCTCCCTGCTGGGAAGGCAGGGACCAAGCCCCAGTACCTATTCATCTCTCTCTAGGTCAGAGCTTCTTCCTC
ACACTGGAGGGGAAGCCTG
GGTCCCGAGTAGCCACGGTCCCTTGTC
mm4__chr17 81.51 Mbp_chr17_81512246 CCATCCACTCACCTGGTGTCTGGGGCGCCCGACGATGGAAGGAAACACGGCTCGGGGAGCGTCGTCCCCAGCAAAACCAG
CTTTGCACATGCCGGAGCCATTGTCAATGACCAGCGCGGCGATCTCTTCTTCCAT
mm3__chr10 71.74 Mbp_chr10_71742343 CAGAAAGGGTGCCTCTCTGGCCCTCTTCCCCTACATTGTGTGGTGTGGCATGCTGTCACAACACGTCAGGGT
mm4__chr1 110.21 Mbp_chr1_110209706 GCCAGAGCTACGGTTTCCTTCCATCCGGCACAAGAAGCGCTTCTCTGCCCAAGGCCACCCCGTCCTCCATTCCATCGCGG
GG
ACTTTGGTGGCTGGGACAGAGGCGGAGAGCATCAAGACCAT
mm4__chr2 241.32 Mbp_chr2_241315785 CAACTGCCGTGGGAGGAGCACGGCGCTTGCGCAGAGGCGGACAACGCGGACGACTCTGGGCTTGCGCACTCGGAAGCCGG
CCGCATGGGGGAGGGGAAGAAGGCCAGGTCGCACTGAGCCAGGAAGCAGTT
mm4__chr2 96.13 Mbp_chr2_96131842 TGAAAGGTCTCGTGCTGGACGCCCTCCACCTGCGGTAGGCCTTCCTGCTCCAACCCCTAAGCCAGACTCCCACCCAAACC
TCCTCTGCCCCTCCCTCGTGGGGCTCTTCCTGCCTCCTGAACCAGACCATCAAGCCCA
mm4__chr15 87.95 Mbp_chr15_87953912 ACACATTGTCTGCAGGCTGTGGCCTCCTTGGGAATAGACCACTCCACTTTCTCTGCTTTCCCTGGGAACGAGTTCTTTCT
AGCCCCCTTGCCCATCTGCCCCTCCTCCGTGCGGGTTATCGACCTCCATTGCCAGC
mm4__chr10 124.53 Mbp_chr10_124526823 CCAGAAGTCCCATCGTTGCTGCCTTTGTGGTTAAAGTCCGCTGAACCCAAGCGCTCTCTTCCCCTCCAGTGTGTGGTGGG
GAGGTGGGGAGGGGACCCAGGCAGATCTGGGAAGCC
mm4__chr19 10.09 Mbp_chr19_10086076 GAGGAGGGTGATGAACTGGGGGAGCGCGCGCTGGGTGGGGGGGCGCGGGGCCTCGCCGCGGCCTTCTGATGCGGCCCCGC
CCCCACGAGGGACGTGAAGACGCGGACCCCGCGCAGCGTGGGGACATGTGACCGACTGCA
mm4__chr1 208.02 Mbp_chr1_208016929 AGGCAGAGGAGAAGTCAGGTACCGTCAGGCTGGGGAGCCTCCCTTCTTACCCTCCAGTGTGTGGCACTGTAGGCTCTTCT
GTGGGG
mm4__chr1 181.48 Mbp_chr1_181481588 GTTTCCTTCCTCAGCCCCTCCTTTCCTTCTTCCCTTCCATTGTTCGGGTTGGAAGGATTTGGAGAGGCCTATGTGTTGAG
CCAAGGCAGGAAAACACAG
mm4__chr17 3.92 Mbp_chr17_3915975 CCTCGTCCCCATCTCAACAGCGAGTTGGCAGGATTTTCTATTCTCGCACAGTGGCTTGCCAGGAAGGCCTTCCCCTGGAT
GGAGAGGAAGAGGG
GCTCGCTGGTGTCACGCAAGGGATGTCAGA
mm4__chr3 143.86 Mbp_chr3_143855297 GGAAAGCCGAGGACAGCATACTCTGAGACACACTTGCTGGGGTAAAATCTATCTGCACAGTAGAATCTACTTCCAAGTAC
TCAACCTCTCTATAGAGGGGAAGAAGGGTTGCCTTTTGCAGAGACAGCAG
mm4__chr1 15.74 Mbp_chr1_15742593 GGGCTGCCATGTTCTCTCTTCCATCCCTCCCCTCCTGGCTCCCCGGCCTCCCCTCCCTCGAGTGGGGCTCTAGCCTCCTT
GACTCCCTCC
mm4__chr22 17.02 Mbp_chr22_17015514 ATGCTCAAAAGGAAGGGCCAGGTTTCCCTGACACAGTGGGCTCCGTGGTCTCCTCCCCTCCTTTTCACCTCAGGAGGGAG
GGAAAGAGGG
GAAGAGAAGGTCCTGCAGAGGA
mm4__chr22 11.02 Mbp_chr22_11017397 TCCTCTGCAGGACCTTCTCTTCCCCTCTTTCCCTCCCTCCTGAGGTGAAAAGGAGGGGAGGAGACCACGGAGCCCTCTGT
GTCAGGGAAACCTGGCCCTTCCTTTTGAGCGTTCAGGGGACTC
mm4__chr21 10.36 Mbp_chr21_10364477 TCCTCTGCAGGACCTTCTCTTCCCCTCTTTCCCTCCCTCCTGAGGTGAAAAGGAGGGGAGGAGACCACGGAGCCCTCTGT
GTCAGGGAAACCTGGCCCTTCCTTTTGAGCGTTCAGGGGACTC
mm4__chr22 50.44 Mbp_chr22_50438048 CTCGGAACAGTCGCTTTCCTTGCCCCTCCCCGTCCTCCCCTCCAGCCCGGGGCTCCCACATCCCGAACTAAGCCCCTCT
mm4__chr10 129.46 Mbp_chr10_129461459 CCTGCCTTTCCTTCTCTGGGCTGTAGTTCACTTTCTAATAAAGGTCCTTTGGGTTTGGCGGCTTTGTTCTGGGCCTCCTT
CACCTGCATCGTGAGG
AGCACCAGCAAAGGCCGCCTGACACTGCCCTCACCAAGAA
mm4__chrX 72.02 Mbp_chrX_72017129 AAACAACCCACCCCAGAAGGGTGCACTCATTCATCATGCCCCTCTATATAGGGGAAGACGGGCTGGGTAGCAGGGCCTAT
TG
mm4__chr19 12.99 Mbp_chr19_12985900 GAGATCTGATGCCAGCTGGGGCACAGAACCTCCTCCCTCCATCTTCCCCTCCATAGGCTGGGTTTGGAGGAGTGGCCTCT
AGATGCAGC
mm4__chr11 117.48 Mbp_chr11_117483287 ATGAACACACATGGGGCACTTATGCCATCATGAGACCTCGTGCCAGGCTGGGGGGTACGCATGACAATGAGACAGGGTCC
TTGCCTCTGGAGGGAGGGGAAGACAGATACCTGCACCGGCAACATGAACTGGAGGA
mm4__chr21 36.49 Mbp_chr21_36493842 GAACACATGTGGCTGGAGGAAGGTGGGATGGATGAGCACACGCTGGGATTCCTCACAGCCTTCATCTCCTCCATGGTGTG
G
AGGGGACCCAGGATCCTCATGTGTTCCCAAAGCAGGTATAGAGTGGCTGGGCA
mm4__chr5 61.39 Mbp_chr5_61390720 TTTAAGACCCCACCAGCTGCAAAAACTGTTCCTGGCATTAAGCTCCTTCTTCCTTTGCAATTCGGTCTTTCTTGAGTGGT
GCCATGAATGCTGTCTTCTCCTCCATGGTCTGGAAGCAGCCATGGCCAAACTT
mm4__chr8 70.40 Mbp_chr8_70402448 GCTCCAGTCCCCGATTCCCGCCCCTCGGTCCCGCAGGCCCCGGAAACCACTAGAGGGAGGGGAAGAGGGAGGGGCCATCT
AACCAGGGACCGAC
mm4__chr19 14.51 Mbp_chr19_14506115 AACAAGCTCCTGTCCCTTGGCCTGTGTGTCCAGGGCCTGCCAGGCGTCTTCCCCTCCACGCTGCGGGCTATGGGAAGTGG
CTCGGTC
mm3__chr1 223.86 Mbp_chr1_223864743 TGACCCATGCTCACCATCACGCCCTGGTGCCTGGGGCGCCCCACCATGGAAGGGAAGATGGCCCAGGGGTCATCATCCCC
TGTGAAGCCAGCCTTGCACATGCCGGAGCTGTTGTCAATGACGAGCG
mm4__chr9 91.48 Mbp_chr9_91484575 TTTTCCTCTACCCTGCTGCCTCCCGTGTTCCCCACCATGCTGAGGGTGTATGGGCATCCTGGATGTGC
mm4__chr2 218.28 Mbp_chr2_218283001 TGGCCTTGCTAGGGAAGAGAGAGTGAGTGCACGGCCCCTCGCCCTCCCCCTTACCCCCACCCTGGAGGGGGAGACAGATG
TTCTGAATCCCAGGCTCTCTGTGAGTTCTACCTGTCTTCCAGGTGCAGTGCTA
mm4__chr11 67.44 Mbp_chr11_67442136 TCGGGTAGGCCTTGTCAATGCGGCCCGTCTGTGGGCACGAGGGGGCAGTCAGTGGGCTCCATCCCTCCCGAAGCCTTGGT
CTTCCCCTCCATGGAGTGG
GAATGACATGCACGTCCCGGCTCACAGGTGGCCCAGATG
mm4__chr3 184.16 Mbp_chr3_184156562 GCCAGTGGAGAGACGTTAGGAAGGGCCTGGGGCCTGGGTGGGATAGCCAGGAAGAGTTTCTTGGTGTTCTCTATCTTCCC
CTCCCTGGTGGGG
TCTTCCCTTCCACCCAGCTGCAGAGATCTGGACAGGCACAGTTTGGA
mm4__chr19 54.14 Mbp_chr19_54141815 TGACTACTCCAGGGTTCCCTGCCCTGCACACGTAGAGTGGGAGCGGCCCGTGGATCCCAGGGAACTGTGCTTTTCATTGT
AGGCCCCCTCCCTGGAGGGGAAGAGGGCAATCTCCGCTGGTATCTCAGAAGTCT
mm4__chr20 62.59 Mbp_chr20_62590954 ATTCTGGTTCCCTGAGCAGCAGCCCGGGGTGTCAGCAAAGTGTGGGGCATGGACCCCGTCCCCTCTTCCCCTGCATGCTG
GGG
TTCCCATGTCCCCAAGTGCATTGC

Input file for Crispresso

Crispresso, written by Luca Pinello, is a software package to quantify the Cas9-induced mutations on off- or on-targets.

Click here to download an amplicon input file for Crispresso. For each off-target, it includes the off-target name, its PCR amplicon and the guide sequence. Keep a copy of this file.

After sequencing, run CRISPRessoPooled. The tool will map the reads to the amplicons and analyse the mutations:
CRISPRessoPooled -r1 Reads1.fastq.gz -r2 Reads2.fastq.gz -f crisporAmplicons_gYvicTzp9e5VPFC9YwLR.txt --name MY_EXPERIMENT

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CRISPR/Cas9 Guide Designer for chordate vertebrate ecdysozoans lophotrochozoans protostomes spongi corals plants butterflies metazoans genomes fruitflies insects nematodes mammals.