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Contents:

PCR primers for off-targets of GCCGTCTTCCCCTCCATCGT 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_5529550_F TCGTCGGCAGCGTCCTCCCCTACCTGGTGCCT 59.9 1.00
ontarget_mm0__chr7 5.53 Mbp_chr7_5529550_R GTCTCGTGGGCTCGGTGGATGATGATATCGCCGCG 60.4 1.00
mm0__chr11 1.80 Mbp_chr11_1803988_F TCGTCGGCAGCGTCCGCCACCCACATAGAAGTCC 60.4 1.00
mm0__chr11 1.80 Mbp_chr11_1803988_R GTCTCGTGGGCTCGGAACACCTCTGGCATGTGCAA 60.4 1.00
mm1__chr19 9.52 Mbp_chr19_9518233_F TCGTCGGCAGCGTCAGTTAACAATGGCTCCGGCA 59.9 0.90
mm1__chr19 9.52 Mbp_chr19_9518233_R GTCTCGTGGGCTCGGGGAGTCCTTCTGACCGATGC 60.1 0.90
mm1__chr5 77.78 Mbp_chr5_77784970_F TCGTCGGCAGCGTCCTCGTCATTGACAACGGCTC 59.2 0.86
mm1__chr5 77.78 Mbp_chr5_77784970_R GTCTCGTGGGCTCGGAATCCTTCTGACCCATGCCC 59.7 0.86
mm1__chr3 175.98 Mbp_chr3_175978472_F TCGTCGGCAGCGTCGAGTCCTTCTGACCCATGCC 60.1 0.71
mm1__chr3 175.98 Mbp_chr3_175978472_R GTCTCGTGGGCTCGGGCAAAGGCTCCTGTATGTGC 59.5 0.71
mm2__chr10 69.02 Mbp_chr10_69022837_F Primer3: not found at this Tm N.d. 0.66
mm2__chr10 69.02 Mbp_chr10_69022837_R Primer3: not found at this Tm N.d. 0.66
mm1__chr6 88.28 Mbp_chr6_88276900_F TCGTCGGCAGCGTCGAGTCCTTCTGACCCATGCC 60.1 0.64
mm1__chr6 88.28 Mbp_chr6_88276900_R GTCTCGTGGGCTCGGTGGATGATGCTATCACCGCG 60.3 0.64
mm3__chr17 11.27 Mbp_chr17_11274628_F TCGTCGGCAGCGTCCGGTCTTTTCCTCATGGGCT 60.0 0.62
mm3__chr17 11.27 Mbp_chr17_11274628_R GTCTCGTGGGCTCGGTGGCACATCTAGCTACGGAT 58.2 0.62
mm3__chr6 46.21 Mbp_chr6_46205059_F TCGTCGGCAGCGTCCGGCATGTGAAAAGCAGCTT 60.0 0.59
mm3__chr6 46.21 Mbp_chr6_46205059_R GTCTCGTGGGCTCGGCACGTAGGAGTCCTTCTGGC 59.8 0.59
mm2__chr19 30.69 Mbp_chr19_30685761_F TCGTCGGCAGCGTCTAAAGAAATGGCCCCGTGCA 60.2 0.49
mm2__chr19 30.69 Mbp_chr19_30685761_R GTCTCGTGGGCTCGGGATCTCAGCAACGTGGCAGG 61.3 0.49
mm2__chr22 41.08 Mbp_chr22_41075132_F TCGTCGGCAGCGTCGGAGTCCTTCTGACCCATGC 60.1 0.48
mm2__chr22 41.08 Mbp_chr22_41075132_R GTCTCGTGGGCTCGGTGTTGACAACAGCTCTGGCA 60.1 0.48
mm2__chr11 50.00 Mbp_chr11_49996391_F Primer3: not found at this Tm N.d. 0.47
mm2__chr11 50.00 Mbp_chr11_49996391_R Primer3: not found at this Tm N.d. 0.47
mm3__chr7 68.00 Mbp_chr7_67998280_F TCGTCGGCAGCGTCTCCAATCCCAACCAAACCGT 59.8 0.45
mm3__chr7 68.00 Mbp_chr7_67998280_R GTCTCGTGGGCTCGGACCTCTCACCTGTAGCCTGA 59.5 0.45
mm2__chr2 73.90 Mbp_chr2_73901398_F TCGTCGGCAGCGTCAGGCTTCGCAGGAGATGATG 59.8 0.44
mm2__chr2 73.90 Mbp_chr2_73901398_R GTCTCGTGGGCTCGGTATCCAGATGAGCACGCACC 59.8 0.44
mm4__chr5 14.27 Mbp_chr5_14268292_F TCGTCGGCAGCGTCTGGCCTGGAATATGACAGCC 59.8 0.43
mm4__chr5 14.27 Mbp_chr5_14268292_R GTCTCGTGGGCTCGGAGAGCTGTGTGACCTGAGTC 59.0 0.43
mm3__chr15 43.99 Mbp_chr15_43990075_F TCGTCGGCAGCGTCGAGTCCTTCTGACCCATGCC 60.1 0.42
mm3__chr15 43.99 Mbp_chr15_43990075_R GTCTCGTGGGCTCGGTGTCGTCGACAATGGCTCTG 60.3 0.42
mm4__chr5 141.64 Mbp_chr5_141644322_F Primer3: not found at this Tm N.d. 0.41
mm4__chr5 141.64 Mbp_chr5_141644322_R Primer3: not found at this Tm N.d. 0.41
mm2__chr5 131.66 Mbp_chr5_131660009_F TCGTCGGCAGCGTCATGCCCATCATCACACCCTG 60.1 0.38
mm2__chr5 131.66 Mbp_chr5_131660009_R GTCTCGTGGGCTCGGTGTGGAAGGTCTGCTTCACG 60.2 0.38
mm4__chr9 36.73 Mbp_chr9_36726254_F TCGTCGGCAGCGTCGGAGAACGTCCTGCCTTCTC 60.1 0.37
mm4__chr9 36.73 Mbp_chr9_36726254_R GTCTCGTGGGCTCGGGACTGGTTCTGAGGCCTAGC 59.8 0.37
mm2__chr22 15.72 Mbp_chr22_15721570_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr22 15.72 Mbp_chr22_15721570_R GTCTCGTGGGCTCGGATCATGTTCTGGTGCCTGGG 60.0 0.37
mm2__chr2 131.26 Mbp_chr2_131263639_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr2 131.26 Mbp_chr2_131263639_R GTCTCGTGGGCTCGGCTCTTTCTGATGCATGCCCC 58.9 0.37
mm2__chr2 130.66 Mbp_chr2_130656833_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr2 130.66 Mbp_chr2_130656833_R GTCTCGTGGGCTCGGCTCTTTCTGATGCATGCCCC 58.9 0.37
mm2__chr2 130.46 Mbp_chr2_130463836_F TCGTCGGCAGCGTCCTCTTTCTGATGCATGCCCC 58.9 0.37
mm2__chr2 130.46 Mbp_chr2_130463836_R GTCTCGTGGGCTCGGGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr2 130.08 Mbp_chr2_130075264_F TCGTCGGCAGCGTCCTCTTTCTGATGCATGCCCC 58.9 0.37
mm2__chr2 130.08 Mbp_chr2_130075264_R GTCTCGTGGGCTCGGGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr14 19.40 Mbp_chr14_19402801_F TCGTCGGCAGCGTCATCATGTTCTGGTGCCTGGG 60.0 0.37
mm2__chr14 19.40 Mbp_chr14_19402801_R GTCTCGTGGGCTCGGGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr14 19.00 Mbp_chr14_18998951_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.37
mm2__chr14 19.00 Mbp_chr14_18998951_R GTCTCGTGGGCTCGGATCATGTTCTGGTGCCTGGG 60.0 0.37
mm4__chr12 56.61 Mbp_chr12_56611571_F TCGTCGGCAGCGTCTCAGAGGAAAAGAGGCATTCCC 60.0 0.34
mm4__chr12 56.61 Mbp_chr12_56611571_R GTCTCGTGGGCTCGGGGAGACGTATTGCTACCCCAG 59.9 0.34
mm3__chr11 48.38 Mbp_chr11_48384229_F Primer3: not found at this Tm N.d. 0.32
mm3__chr11 48.38 Mbp_chr11_48384229_R Primer3: not found at this Tm N.d. 0.32
mm4__chr14 50.12 Mbp_chr14_50117696_F Primer3: not found at this Tm N.d. 0.30
mm4__chr14 50.12 Mbp_chr14_50117696_R Primer3: not found at this Tm N.d. 0.30
mm2__chr15 34.79 Mbp_chr15_34794695_F TCGTCGGCAGCGTCGGGAAGTTTACCTGGTGCCG 60.9 0.30
mm2__chr15 34.79 Mbp_chr15_34794695_R GTCTCGTGGGCTCGGATGTGTGACGACGAGGAGAC 59.4 0.30
mm4__chr6 41.54 Mbp_chr6_41537183_F Primer3: not found at this Tm N.d. 0.29
mm4__chr6 41.54 Mbp_chr6_41537183_R Primer3: not found at this Tm N.d. 0.29
mm4__chr9 102.38 Mbp_chr9_102383949_F Primer3: not found at this Tm N.d. 0.29
mm4__chr9 102.38 Mbp_chr9_102383949_R Primer3: not found at this Tm N.d. 0.29
mm4__chr10 88.95 Mbp_chr10_88948817_F TCGTCGGCAGCGTCCTCACCTGATGTCTGGGACG 59.8 0.29
mm4__chr10 88.95 Mbp_chr10_88948817_R GTCTCGTGGGCTCGGACTGCCTTGGTGTGTGACAA 60.0 0.29
mm3__chr6 101.43 Mbp_chr6_101431430_F TCGTCGGCAGCGTCTGATGCTGTCCTGTGTGCTT 59.8 0.28
mm3__chr6 101.43 Mbp_chr6_101431430_R GTCTCGTGGGCTCGGCGGCTTTGCTGGGGATGA 60.0 0.28
mm4__chrY 18.15 Mbp_chrY_18147967_F TCGTCGGCAGCGTCACAATGGCTCTTGCATGTGC 59.7 0.27
mm4__chrY 18.15 Mbp_chrY_18147967_R GTCTCGTGGGCTCGGGAGTCCTTCTGGCCCATGTC 60.1 0.27
mm4__chrY 17.76 Mbp_chrY_17758018_F TCGTCGGCAGCGTCGAGTCCTTCTGGCCCATGTC 60.1 0.27
mm4__chrY 17.76 Mbp_chrY_17758018_R GTCTCGTGGGCTCGGACAATGGCTCTTGCATGTGC 59.7 0.27
mm3__chr8 72.27 Mbp_chr8_72268161_F Primer3: not found at this Tm N.d. 0.27
mm3__chr8 72.27 Mbp_chr8_72268161_R Primer3: not found at this Tm N.d. 0.27
mm3__chr11 36.67 Mbp_chr11_36666775_F Primer3: not found at this Tm N.d. 0.27
mm3__chr11 36.67 Mbp_chr11_36666775_R Primer3: not found at this Tm N.d. 0.27
mm4__chr16 5.51 Mbp_chr16_5508661_F TCGTCGGCAGCGTCAGAGTGCCACACCCAAGATG 59.9 0.27
mm4__chr16 5.51 Mbp_chr16_5508661_R GTCTCGTGGGCTCGGACTCTGTGCAATCGCTCTGT 59.6 0.27
mm4__chr19 14.20 Mbp_chr19_14197725_F Primer3: not found at this Tm N.d. 0.26
mm4__chr19 14.20 Mbp_chr19_14197725_R Primer3: not found at this Tm N.d. 0.26
mm4__chr3 58.55 Mbp_chr3_58552641_F Primer3: not found at this Tm N.d. 0.26
mm4__chr3 58.55 Mbp_chr3_58552641_R Primer3: not found at this Tm N.d. 0.26
mm4__chr17 47.39 Mbp_chr17_47388227_F Primer3: not found at this Tm N.d. 0.26
mm4__chr17 47.39 Mbp_chr17_47388227_R Primer3: not found at this Tm N.d. 0.26
mm4__chr16 5.91 Mbp_chr16_5905904_F TCGTCGGCAGCGTCCACTCCCCAAATGCCTCCTC 60.3 0.26
mm4__chr16 5.91 Mbp_chr16_5905904_R GTCTCGTGGGCTCGGCCTAAGGTAGGGCTTTGGGC 60.1 0.26
mm4__chr16 50.06 Mbp_chr16_50059124_F TCGTCGGCAGCGTCCGCCCACATAGGAGTCCTTC 59.8 0.26
mm4__chr16 50.06 Mbp_chr16_50059124_R GTCTCGTGGGCTCGGTGGCTTTGCTAGGGACGATG 60.1 0.26
mm4__chr1 186.61 Mbp_chr1_186606328_F Primer3: not found at this Tm N.d. 0.25
mm4__chr1 186.61 Mbp_chr1_186606328_R Primer3: not found at this Tm N.d. 0.25
mm4__chr22 45.67 Mbp_chr22_45674071_F Primer3: not found at this Tm N.d. 0.25
mm4__chr22 45.67 Mbp_chr22_45674071_R Primer3: not found at this Tm N.d. 0.25
mm3__chr17 81.51 Mbp_chr17_81512247_F TCGTCGGCAGCGTCCCATCCACTCACCTGGTGTC 60.0 0.23
mm3__chr17 81.51 Mbp_chr17_81512247_R GTCTCGTGGGCTCGGATGGAAGAAGAGATCGCCGC 60.2 0.23
mm4__chr7 63.40 Mbp_chr7_63404892_F TCGTCGGCAGCGTCAGGAAACTCATCAGCTTGGGT 59.2 0.22
mm4__chr7 63.40 Mbp_chr7_63404892_R GTCTCGTGGGCTCGGTTCCAGCAAAAGGAGTGGCT 59.8 0.22
mm4__chr7 63.34 Mbp_chr7_63343195_F TCGTCGGCAGCGTCTTCCAGCAAAAGGAGTGGCT 59.8 0.22
mm4__chr7 63.34 Mbp_chr7_63343195_R GTCTCGTGGGCTCGGAGGAAACTCATCAGCTTGGGT 59.2 0.22
mm2__chr1 229.43 Mbp_chr1_229433002_F TCGTCGGCAGCGTCGCAGCCTGACCTGGTGTC 60.3 0.22
mm2__chr1 229.43 Mbp_chr1_229433002_R GTCTCGTGGGCTCGGGACGAAGACGAGACCACCG 60.1 0.22
mm2__chr18 62.44 Mbp_chr18_62442092_F TCGTCGGCAGCGTCCCTCGTCATCGACAATGGCT 60.1 0.22
mm2__chr18 62.44 Mbp_chr18_62442092_R GTCTCGTGGGCTCGGGAGTCCTTCTGACCCATGCC 60.1 0.22
mm4__chr1 237.93 Mbp_chr1_237926914_F TCGTCGGCAGCGTCTGACAATGGCTCCAGCATGT 59.9 0.21
mm4__chr1 237.93 Mbp_chr1_237926914_R GTCTCGTGGGCTCGGCACGTAGGAGTCCTTCTGGC 59.8 0.21
mm4__chr12 26.33 Mbp_chr12_26334148_F TCGTCGGCAGCGTCCCAAGATCACAAAGATACACCTG 57.1 0.20
mm4__chr12 26.33 Mbp_chr12_26334148_R GTCTCGTGGGCTCGGCCCTGCTCTAATGACTGTGCT 59.7 0.20
mm4__chr10 85.62 Mbp_chr10_85617054_F TCGTCGGCAGCGTCATCCTGTCCTCTGCCAAAGC 60.0 0.20
mm4__chr10 85.62 Mbp_chr10_85617054_R GTCTCGTGGGCTCGGCAGGTCCCAGCCTTATGTGG 60.1 0.20
mm4__chrX 86.32 Mbp_chrX_86322420_F Primer3: not found at this Tm N.d. 0.19
mm4__chrX 86.32 Mbp_chrX_86322420_R Primer3: not found at this Tm N.d. 0.19
mm4__chr8 43.43 Mbp_chr8_43431851_F Primer3: not found at this Tm N.d. 0.19
mm4__chr8 43.43 Mbp_chr8_43431851_R Primer3: not found at this Tm N.d. 0.19
mm4__chr12 60.65 Mbp_chr12_60648400_F Primer3: not found at this Tm N.d. 0.19
mm4__chr12 60.65 Mbp_chr12_60648400_R Primer3: not found at this Tm N.d. 0.19
mm4__chr11 114.59 Mbp_chr11_114587686_F Primer3: not found at this Tm N.d. 0.19
mm4__chr11 114.59 Mbp_chr11_114587686_R Primer3: not found at this Tm N.d. 0.19
mm4__chr10 95.13 Mbp_chr10_95131523_F Primer3: not found at this Tm N.d. 0.19
mm4__chr10 95.13 Mbp_chr10_95131523_R Primer3: not found at this Tm N.d. 0.19
mm4__chr1 42.51 Mbp_chr1_42511089_F Primer3: not found at this Tm N.d. 0.19
mm4__chr1 42.51 Mbp_chr1_42511089_R Primer3: not found at this Tm N.d. 0.19
mm3__chr20 57.92 Mbp_chr20_57918621_F Primer3: not found at this Tm N.d. 0.18
mm3__chr20 57.92 Mbp_chr20_57918621_R Primer3: not found at this Tm N.d. 0.18
mm4__chr17 39.08 Mbp_chr17_39083400_F Primer3: not found at this Tm N.d. 0.18
mm4__chr17 39.08 Mbp_chr17_39083400_R Primer3: not found at this Tm N.d. 0.18
mm4__chr1 75.91 Mbp_chr1_75909149_F Primer3: not found at this Tm N.d. 0.17
mm4__chr1 75.91 Mbp_chr1_75909149_R Primer3: not found at this Tm N.d. 0.17
mm4__chr2 129.76 Mbp_chr2_129764606_F Primer3: not found at this Tm N.d. 0.17
mm4__chr2 129.76 Mbp_chr2_129764606_R Primer3: not found at this Tm N.d. 0.17
mm4__chr11 54.56 Mbp_chr11_54558483_F Primer3: not found at this Tm N.d. 0.17
mm4__chr11 54.56 Mbp_chr11_54558483_R Primer3: not found at this Tm N.d. 0.17
mm4__chr3 38.55 Mbp_chr3_38554277_F TCGTCGGCAGCGTCTTCTCCGTCCAGCTGACTTG 59.6 0.16
mm4__chr3 38.55 Mbp_chr3_38554277_R GTCTCGTGGGCTCGGCCCTGCGCCACTACTACTTC 60.1 0.16
mm4__chr19 49.37 Mbp_chr19_49374719_F TCGTCGGCAGCGTCCCTTGTTCTTCCTCCCCAGG 59.6 0.15
mm4__chr19 49.37 Mbp_chr19_49374719_R GTCTCGTGGGCTCGGGGGGATGGAGTTCTGTGTGG 60.0 0.15
mm4__chr21 32.00 Mbp_chr21_31998305_F TCGTCGGCAGCGTCAAGCAGGGACGATGTTAGGC 60.1 0.15
mm4__chr21 32.00 Mbp_chr21_31998305_R GTCTCGTGGGCTCGGACCAGCAGCATGTCACTCTC 60.0 0.15
mm4__chr7 65.40 Mbp_chr7_65404303_F Primer3: not found at this Tm N.d. 0.14
mm4__chr7 65.40 Mbp_chr7_65404303_R Primer3: not found at this Tm N.d. 0.14
mm4__chr19 22.88 Mbp_chr19_22883631_F Primer3: not found at this Tm N.d. 0.14
mm4__chr19 22.88 Mbp_chr19_22883631_R Primer3: not found at this Tm N.d. 0.14
mm4__chr19 2.08 Mbp_chr19_2078138_F Primer3: not found at this Tm N.d. 0.14
mm4__chr19 2.08 Mbp_chr19_2078138_R Primer3: not found at this Tm N.d. 0.14
mm4__chr12 126.88 Mbp_chr12_126884173_F TCGTCGGCAGCGTCCAGGAGTCACAGAGTGGGGA 60.5 0.14
mm4__chr12 126.88 Mbp_chr12_126884173_R GTCTCGTGGGCTCGGCCAGAATAACTCTTGCTTTGGGG 59.5 0.14
mm4__chr1 31.16 Mbp_chr1_31161664_F Primer3: not found at this Tm N.d. 0.13
mm4__chr1 31.16 Mbp_chr1_31161664_R Primer3: not found at this Tm N.d. 0.13
mm4__chr19 40.40 Mbp_chr19_40398533_F TCGTCGGCAGCGTCTGACAAGACAGAGGGCAAGG 59.6 0.12
mm4__chr19 40.40 Mbp_chr19_40398533_R GTCTCGTGGGCTCGGAAGCTGGTACGCGTGCTTAG 60.7 0.12
mm4__chr16 48.58 Mbp_chr16_48578495_F TCGTCGGCAGCGTCGTTATCCCACACGTGCCTCA 60.0 0.12
mm4__chr16 48.58 Mbp_chr16_48578495_R GTCTCGTGGGCTCGGTCACAAGACTCAGTGCCCAC 59.8 0.12
mm4__chr15_KI270727v1_random 283.41 Kbp_chr15_KI270727v1_random_283406_F Primer3: not found at this Tm N.d. 0.12
mm4__chr15_KI270727v1_random 283.41 Kbp_chr15_KI270727v1_random_283406_R Primer3: not found at this Tm N.d. 0.12
mm4__chr15 21.99 Mbp_chr15_21993604_F Primer3: not found at this Tm N.d. 0.12
mm4__chr15 21.99 Mbp_chr15_21993604_R Primer3: not found at this Tm N.d. 0.12
mm4__chr15 21.56 Mbp_chr15_21556311_F Primer3: not found at this Tm N.d. 0.12
mm4__chr15 21.56 Mbp_chr15_21556311_R Primer3: not found at this Tm N.d. 0.12
mm4__chr15 20.98 Mbp_chr15_20982572_F Primer3: not found at this Tm N.d. 0.12
mm4__chr15 20.98 Mbp_chr15_20982572_R Primer3: not found at this Tm N.d. 0.12
mm4__chr4 689.11 Kbp_chr4_689110_F Primer3: not found at this Tm N.d. 0.11
mm4__chr4 689.11 Kbp_chr4_689110_R Primer3: not found at this Tm N.d. 0.11
mm4__chr10 129.19 Mbp_chr10_129192606_F TCGTCGGCAGCGTCTGGTGACAGCACAGCAGTAG 59.9 0.10
mm4__chr10 129.19 Mbp_chr10_129192606_R GTCTCGTGGGCTCGGCTGGCAAGATCCAATCTGCC 58.9 0.10
mm4__chr1 112.00 Mbp_chr1_112004369_F TCGTCGGCAGCGTCTCTTTCTGCCCATAGCCTGG 59.4 0.10
mm4__chr1 112.00 Mbp_chr1_112004369_R GTCTCGTGGGCTCGGTGTTTGCCTCTGAGGAAGACC 59.9 0.10
mm4__chr17 79.10 Mbp_chr17_79104967_F TCGTCGGCAGCGTCACTGTTCAGGTCGTTTGGGC 60.8 0.09
mm4__chr17 79.10 Mbp_chr17_79104967_R GTCTCGTGGGCTCGGCAGTGAGAGAGACAGGGCTG 59.4 0.09
mm3__chr5 147.47 Mbp_chr5_147466725_F TCGTCGGCAGCGTCAAGCCAACCCCAAAGTGTGA 60.0 0.09
mm3__chr5 147.47 Mbp_chr5_147466725_R GTCTCGTGGGCTCGGACAACACAGCCAAAGTCCCT 59.7 0.09
mm4__chr1 61.74 Mbp_chr1_61743012_F TCGTCGGCAGCGTCTATTTTGCGCCCAGATCGGT 60.1 0.09
mm4__chr1 61.74 Mbp_chr1_61743012_R GTCTCGTGGGCTCGGCCCTCCTCTTTGTCCCCTCA 60.5 0.09
mm4__chr11 2.41 Mbp_chr11_2412146_F TCGTCGGCAGCGTCACCTGAGTGGCTTCATCTGG 59.3 0.09
mm4__chr11 2.41 Mbp_chr11_2412146_R GTCTCGTGGGCTCGGGGGGACAACTGGAACAAGTG 58.6 0.09
mm4__chr7 75.81 Mbp_chr7_75810382_F Primer3: not found at this Tm N.d. 0.07
mm4__chr7 75.81 Mbp_chr7_75810382_R Primer3: not found at this Tm N.d. 0.07
mm3__chr1 246.53 Mbp_chr1_246530782_F Primer3: not found at this Tm N.d. 0.07
mm3__chr1 246.53 Mbp_chr1_246530782_R Primer3: not found at this Tm N.d. 0.07
mm4__chr13 112.88 Mbp_chr13_112883403_F TCGTCGGCAGCGTCTGCCTTTCAGGAAAGCACCA 60.1 0.07
mm4__chr13 112.88 Mbp_chr13_112883403_R GTCTCGTGGGCTCGGCACCTCGGAAACACCCTCTG 60.3 0.07
mm3__chr1 223.86 Mbp_chr1_223864744_F TCGTCGGCAGCGTCTGACCCATGCTCACCATCAC 60.0 0.06
mm3__chr1 223.86 Mbp_chr1_223864744_R GTCTCGTGGGCTCGGCGCTCGTCATTGACAACAGC 60.1 0.06
mm4__chr11 111.28 Mbp_chr11_111284161_F TCGTCGGCAGCGTCGGTCACGTCAGGTTACTACGG 60.1 0.05
mm4__chr11 111.28 Mbp_chr11_111284161_R GTCTCGTGGGCTCGGGAGGACTCACAGGGAAACGG 60.0 0.05
mm2__chr3 12.07 Mbp_chr3_12070308_F TCGTCGGCAGCGTCTCATTGACAGTGTCTCCGGC 60.0 0.05
mm2__chr3 12.07 Mbp_chr3_12070308_R GTCTCGTGGGCTCGGGAGTCCTTCTGACCCATGCC 60.1 0.05
mm4__chr4 189.86 Mbp_chr4_189862364_F Primer3: not found at this Tm N.d. 0.05
mm4__chr4 189.86 Mbp_chr4_189862364_R Primer3: not found at this Tm N.d. 0.05
mm3__chr12 89.12 Mbp_chr12_89123197_F TCGTCGGCAGCGTCACCAGATGAATATGTTAAATGCACC 57.9 0.05
mm3__chr12 89.12 Mbp_chr12_89123197_R GTCTCGTGGGCTCGGGCATGTACACCCAAGAAAAGCA 59.7 0.05
mm4__chr16 49.52 Mbp_chr16_49524459_F TCGTCGGCAGCGTCAGTGTGGCAGTGACAGATGG 59.9 0.04
mm4__chr16 49.52 Mbp_chr16_49524459_R GTCTCGTGGGCTCGGTGTCCACTCTGGGGCAAATC 59.9 0.04
mm4__chr3 76.49 Mbp_chr3_76486037_F Primer3: not found at this Tm N.d. 0.04
mm4__chr3 76.49 Mbp_chr3_76486037_R Primer3: not found at this Tm N.d. 0.04
mm4__chr20 50.24 Mbp_chr20_50243254_F TCGTCGGCAGCGTCGGCACAAAGGCCCATTGTTC 60.3 0.04
mm4__chr20 50.24 Mbp_chr20_50243254_R GTCTCGTGGGCTCGGGTGAATCGCTCCTCAGTCCC 60.1 0.04
mm4__chr2 27.37 Mbp_chr2_27373994_F TCGTCGGCAGCGTCAGGATGTCCTGGAGGTGAGG 60.3 0.04
mm4__chr2 27.37 Mbp_chr2_27373994_R GTCTCGTGGGCTCGGCAGCTACAGCCTGGGGATAG 59.3 0.04
mm4__chr18 58.26 Mbp_chr18_58255782_F TCGTCGGCAGCGTCTTCATGGCATTCCACACCCT 59.5 0.03
mm4__chr18 58.26 Mbp_chr18_58255782_R GTCTCGTGGGCTCGGATGGACGAGCTCCGCTTAAG 59.8 0.03
mm4__chr9 93.95 Mbp_chr9_93947410_F TCGTCGGCAGCGTCTCCCCTGTGAATTCCTTGGC 59.9 0.03
mm4__chr9 93.95 Mbp_chr9_93947410_R GTCTCGTGGGCTCGGCCACTCCCATGGGTGTACTG 59.7 0.03
mm4__chr5 57.48 Mbp_chr5_57482597_F TCGTCGGCAGCGTCATCTCCCACGTAGCAGTCCT 60.0 0.03
mm4__chr5 57.48 Mbp_chr5_57482597_R GTCTCGTGGGCTCGGATGGGTCAGGGATGTGCAAG 60.0 0.03
mm4__chr8 84.95 Mbp_chr8_84948672_F TCGTCGGCAGCGTCCTCTGGTATGTGCAAGGCCA 60.0 0.03
mm4__chr8 84.95 Mbp_chr8_84948672_R GTCTCGTGGGCTCGGCAGTCCTTCTGCCCTATGCC 60.1 0.03
mm4__chr10 122.46 Mbp_chr10_122459473_F Primer3: not found at this Tm N.d. 0.03
mm4__chr10 122.46 Mbp_chr10_122459473_R Primer3: not found at this Tm N.d. 0.03
mm4__chr1 202.87 Mbp_chr1_202873375_F TCGTCGGCAGCGTCTGCTGGGTATAGTTCCCCGA 60.0 0.02
mm4__chr1 202.87 Mbp_chr1_202873375_R GTCTCGTGGGCTCGGGGAGTCCTTCTGGCCTATGC 59.8 0.02
mm4__chr19 3.99 Mbp_chr19_3985433_F TCGTCGGCAGCGTCGAACCGAAAGAAGCGAGTCG 59.2 0.02
mm4__chr19 3.99 Mbp_chr19_3985433_R GTCTCGTGGGCTCGGAGCTGAGTGTTGACGTCAGC 60.5 0.02
mm3__chr2 131.63 Mbp_chr2_131626222_F TCGTCGGCAGCGTCGTGCTCGTCATTGACAACGG 59.8 0.02
mm3__chr2 131.63 Mbp_chr2_131626222_R GTCTCGTGGGCTCGGACTCCTTCTGATGCATGCCC 60.1 0.02
mm4__chr20 57.73 Mbp_chr20_57734279_F TCGTCGGCAGCGTCGCTGCCCCATCTCAAGGAAT 60.1 0.02
mm4__chr20 57.73 Mbp_chr20_57734279_R GTCTCGTGGGCTCGGGAGACCAGGTGTGCAAAAGC 59.6 0.02
mm4__chr11 112.53 Mbp_chr11_112529298_F TCGTCGGCAGCGTCTGAGGAGGGTAGCACCAGAA 59.8 0.02
mm4__chr11 112.53 Mbp_chr11_112529298_R GTCTCGTGGGCTCGGTGTACAATCACGGGCCACTC 60.0 0.02
mm4__chr5 5.73 Mbp_chr5_5725799_F TCGTCGGCAGCGTCATCTGGGGCATGGAAATGGG 60.1 0.02
mm4__chr5 5.73 Mbp_chr5_5725799_R GTCTCGTGGGCTCGGTGCTGCTCTTGAAAAGGCAG 59.0 0.02
mm4__chr3 157.81 Mbp_chr3_157807133_F Primer3: not found at this Tm N.d. 0.01
mm4__chr3 157.81 Mbp_chr3_157807133_R Primer3: not found at this Tm N.d. 0.01
mm4__chr5 98.34 Mbp_chr5_98338826_F TCGTCGGCAGCGTCGCTCCTCATCAACAAGGGCT 60.0 0.00
mm4__chr5 98.34 Mbp_chr5_98338826_R GTCTCGTGGGCTCGGTTTTGACCCATGCCCACCAT 60.1 0.00
mm4__chr16 2.52 Mbp_chr16_2523272_F TCGTCGGCAGCGTCCTAGTTCTGTCCAGAGCCCC 59.1 0.00
mm4__chr16 2.52 Mbp_chr16_2523272_R GTCTCGTGGGCTCGGCACACACCAAGAGAGGGACC 59.9 0.00
mm4__chr19 54.14 Mbp_chr19_54141816_F TCGTCGGCAGCGTCTGACTACTCCAGGGTTCCCT 59.2 0.00
mm4__chr19 54.14 Mbp_chr19_54141816_R GTCTCGTGGGCTCGGAGACTTCTGAGATACCAGCGG 58.9 0.00
mm4__chr3 39.18 Mbp_chr3_39180374_F Primer3: not found at this Tm N.d. 0.00
mm4__chr3 39.18 Mbp_chr3_39180374_R Primer3: not found at this Tm N.d. 0.00
mm4__chr20 44.31 Mbp_chr20_44313913_F Primer3: not found at this Tm N.d. 0.00
mm4__chr20 44.31 Mbp_chr20_44313913_R Primer3: not found at this Tm N.d. 0.00
mm4__chr2 167.41 Mbp_chr2_167406576_F Primer3: not found at this Tm N.d. 0.00
mm4__chr2 167.41 Mbp_chr2_167406576_R Primer3: not found at this Tm N.d. 0.00
mm4__chr19 33.41 Mbp_chr19_33406847_F TCGTCGGCAGCGTCCCATGTGAGGCCAAGGTGG 60.6 0.00
mm4__chr19 33.41 Mbp_chr19_33406847_R GTCTCGTGGGCTCGGCAGCCCTCACACTCAGCAG 60.3 0.00
mm4__chr16 87.44 Mbp_chr16_87436982_F TCGTCGGCAGCGTCTCTGCACGCAGACACTCATT 59.9 0.00
mm4__chr16 87.44 Mbp_chr16_87436982_R GTCTCGTGGGCTCGGCTTACGCTCCACAGAAGGCA 60.0 0.00
mm4__chr14 69.15 Mbp_chr14_69151684_F Primer3: not found at this Tm N.d. 0.00
mm4__chr14 69.15 Mbp_chr14_69151684_R Primer3: not found at this Tm N.d. 0.00
mm4__chr11 2.27 Mbp_chr11_2269483_F TCGTCGGCAGCGTCGATCCGCCAGCACTAGAGAC 59.9 0.00
mm4__chr11 2.27 Mbp_chr11_2269483_R GTCTCGTGGGCTCGGCAAGACCGGCGTTTGTTTGT 59.9 0.00
mm4__chr11 123.75 Mbp_chr11_123754201_F Primer3: not found at this Tm N.d. 0.00
mm4__chr11 123.75 Mbp_chr11_123754201_R Primer3: not found at this Tm N.d. 0.00
mm4__chr1 46.13 Mbp_chr1_46133074_F TCGTCGGCAGCGTCGCGTCTTTTGTCTGCCTTGC 60.6 0.00
mm4__chr1 46.13 Mbp_chr1_46133074_R GTCTCGTGGGCTCGGTCTGCCTTTGTCCCGCTTAG 60.0 0.00
mm4__chr1 215.08 Mbp_chr1_215081843_F TCGTCGGCAGCGTCTCGCCTATTACGACGACAGC 59.9 0.00
mm4__chr1 215.08 Mbp_chr1_215081843_R GTCTCGTGGGCTCGGCGTAGGGACAGCCAAACTCG 60.7 0.00
mm4__chr1 1.98 Mbp_chr1_1977389_F TCGTCGGCAGCGTCCCCTCTGCATATCCGTCCAC 59.9 0.00
mm4__chr1 1.98 Mbp_chr1_1977389_R GTCTCGTGGGCTCGGCATCTGGGCCAGAGAGGATG 59.6 0.00
mm4__chr1 1.60 Mbp_chr1_1600886_F TCGTCGGCAGCGTCCAGGGCTGGACAAAGGGAAT 59.9 0.00
mm4__chr1 1.60 Mbp_chr1_1600886_R GTCTCGTGGGCTCGGGTGATCTATGCAGGGACGGG 59.9 0.00
mm3__chr18 65.75 Mbp_chr18_65750734_F Primer3: not found at this Tm N.d. 0.00
mm3__chr18 65.75 Mbp_chr18_65750734_R Primer3: not found at this Tm N.d. 0.00
mm3__chr10 116.21 Mbp_chr10_116214403_F Primer3: not found at this Tm N.d. 0.00
mm3__chr10 116.21 Mbp_chr10_116214403_R Primer3: not found at this Tm N.d. 0.00

Off-target amplicon sequences

Primers underlined, off-targets in bold.

ontarget_mm0__chr7 5.53 Mbp_chr7_5529550 CTCCCCTACCTGGTGCCTGGGGCGCCCCACGATGGAGGGGAAGACGGCCCGGGGGGCATCGTCGCCCGCGAAGCCGGCCT
TGCACATGCCGGAGCCGTTGTCGACGACGAGCGCGGCGATATCATCATCCA
mm0__chr11 1.80 Mbp_chr11_1803988 CGCCACCCACATAGAAGTCCTTCTGACCCATGCCCACCGTCATGCCCTGGTGTCTGGGGCACCCCACGATGGAGGGGAAG
ACGGCCCAGGGGGCATCGTCGTCGGTGAAGCTGGCCTTGCACATGCCAGAGGTGTT
mm1__chr19 9.52 Mbp_chr19_9518233 AGTTAACAATGGCTCCGGCATGTGCAAGGTCAACTTCGTGGGCGACAATGCCCCCCGGGCCATCTTCCCCTCCATCGTGG
GGCATCTGAAGCACCAGGGCGTTATGGTGGGCATCGGTCAGAAGGACTCC
mm1__chr5 77.78 Mbp_chr5_77784970 CTCGTCATTGACAACGGCTCCGGCATGTGCAAGGCCAGCTTCACGGGCGACAATGCCGCCCGGGCAGTCTTCCCCTCCAT
CGTTGGGCACCCCAGGCACCAGGGCGTGATGGTGGGCATGGGTCAGAAGGATT
mm1__chr3 175.98 Mbp_chr3_175978472 GAGTCCTTCTGACCCATGCCCACCATCACACCTTGGTGCCTGGGGCGCCCCACGATGGATGGGAAGACGGCCCCGGAGGC
ATCGTCGCCTGCGAAGCTGGTCTTGCACATACAGGAGCCTTTGC
mm1__chr6 88.28 Mbp_chr6_88276900 GAGTCCTTCTGACCCATGCCCACCATCACTCCCTGGTGCCTAGGGTGCCCCACAATGGAGGGGAAGACGGCCTGGGGAGC
CTTGCGCATGCTGGAGCAGTTGTCGACGACGACGAGCGCGGTGATAGCATCATCCA
mm3__chr17 11.27 Mbp_chr17_11274628 CGGTCTTTTCCTCATGGGCTTAATGTCCGTTGCCTACAGAGTAAAGGCTATTTGGGAAACCAAAGATGGAGGAGGAGACG
GCATCCGTAGCTAGATGTGCCA
mm3__chr6 46.21 Mbp_chr6_46205059 CGGCATGTGAAAAGCAGCTTTGCTGGGACAATGCCCTCCGAGCCATATTCCCCTCCATCATCGGGCACCCCCGGCACCAG
GGCGTGATGGTGGGCATGGGCCAGAAGGACTCCTACGTG
mm2__chr19 30.69 Mbp_chr19_30685761 TAAAGAAATGGCCCCGTGCAGAGGCAACCAGGGGGATGGGGGCACCGTCCCAGCCGCTGCCGCCGTCCTCCCCTCCATCA
TGGGCTGGGACGCTGTCAGCTCCACCACCTGCCCCACGGTCCTGCCACGTTGCTGAGATC
mm2__chr22 41.08 Mbp_chr22_41075132 GGAGTCCTTCTGACCCATGCCCACCATCATGACCTGGTGCCTGGTGCACCCCACGATGGAAGGGAAGATGGCCCAGGGGA
CATCGTCACCCGCGAAGCCGGCCTTGCACATGCCAGAGCTGTTGTCAACA
mm3__chr7 68.00 Mbp_chr7_67998280 TCCAATCCCAACCAAACCGTAGAAGTGTCCAAAGATGGAGGGGAAGACAGAGAGGAAGTGGGAAATAGCAGATCAGGCTA
CAGGTGAGAGGT
mm2__chr2 73.90 Mbp_chr2_73901398 AGGCTTCGCAGGAGATGATGCCCCCCGGGCTGTCTTCCCCTCCATTGTGGGCCGCCCTCGCCACCAGGTGCGTGCTCATC
TGGATA
mm4__chr5 14.27 Mbp_chr5_14268292 TGGCCTGGAATATGACAGCCCTCACTTCCATCTTCCACTCCATTGTGGGTATCAGTAAATAGGTGTGATTCAGAAAGTTA
GCAGTAATTGTCAAAAATGCTTTGGAATGATGACTCAGGTCACACAGCTCT
mm3__chr15 43.99 Mbp_chr15_43990075 GAGTCCTTCTGACCCATGCCCACCGTCACGCCCTGGTGTCTGGGGCGCCCCACAATGAAGGGGAAGATGGCCCGGGGTGG
GCATCGTCACCTGCAAAGCTGGCCTTGCACATGCCAGAGCCATTGTCGACGACA
mm2__chr5 131.66 Mbp_chr5_131660009 ATGCCCATCATCACACCCTGGTGCCTCGGGTGCCCCATTATGGAGGGGAAGACGGCCCGGGGGCATCGTTGCCCGTGAAG
CAGACCTTCCACA
mm4__chr9 36.73 Mbp_chr9_36726254 GGAGAACGTCCTGCCTTCTCCATCTTTGTTCCTACTAGTTTTTCACAAAGGGACACATGCAGTTTTCCCCTTCATTGTGG
GTTCCCTATGTGGGAAAATTCCATTTGCTAGGCCTCAGAACCAGTC
mm2__chr22 15.72 Mbp_chr22_15721570 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCAGGTGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGGTGCCCCAGGCACCAGAACATGAT
mm2__chr2 131.26 Mbp_chr2_131263639 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCGGGCGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGGCGCCCCAGGCAGCAGGGCATGATGGGGGGCATGCATCAGAAAGAG
mm2__chr2 130.66 Mbp_chr2_130656833 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCGGGCGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGGTGCCCCAGGCAGCAGGGCATGATGGGGGGCATGCATCAGAAAGAG
mm2__chr2 130.46 Mbp_chr2_130463836 CTCTTTCTGATGCATGCCCCCCATCATGCCCTGCTGCCTGGGGCGCCCCACGATGGAAGGGAAGACAGCCCGGGGGGCAT
CGTCGCCCGCAAAGCCGGCCTTGCACATGCCAGAGCCGTTGTCAATGACGAGCAC
mm2__chr2 130.08 Mbp_chr2_130075264 CTCTTTCTGATGCATGCCCCCCATCATGCCCTGCTGCCTGGGGCGCCCCACGATGGAAGGGAAGACAGCCCGGGGGGCAT
CGTCGCCCGCAAAGCCGGCCTTGCACATGCCAGAGCCGTTGTCAATGACGAGCAC
mm2__chr14 19.40 Mbp_chr14_19402801 ATCATGTTCTGGTGCCTGGGGCACCCCACGATGGAAGGGAAGACAGCCCGGGGGGCATCGTCACCTGCAAAGCCGGCCTT
GCACATGCCAGAGCCGTTGTCAATGACGAGCAC
mm2__chr14 19.00 Mbp_chr14_18998951 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCAGGTGACGATGCCCCCCGGGCTGTCTTCCCTTC
CATCGTGGGGTGCCCCAGGCACCAGAACATGAT
mm4__chr12 56.61 Mbp_chr12_56611571 TCAGAGGAAAAGAGGCATTCCCTTCTTTCTAGAGTTGTGCATTAAACTTGTCAAGGTCAATAAATGTAGCAAACTAGAGC
ATTTACCCATGTTGGAGGGGAAGACGAACTTCTTCTGGGGTAGCAATACGTCTCC
mm2__chr15 34.79 Mbp_chr15_34794695 GGGAAGTTTACCTGGTGCCGCGGGCGGCCCACGATGGACGGGAAGACAGCGCGGGGCGCGTCATCGCCCGCAAAGCCGGC
CTTCACCAGCCCAGAGCCGTTGTCGCACACCAGGGCGGTGGTCTCCTCGTCGTCACACAT
mm4__chr10 88.95 Mbp_chr10_88948817 CTCACCTGATGTCTGGGACGTCCCACAATGGATGGGAAAACAGCCCTGGGAGCATCGTCCCCAGCAAAGCCGGCCTTACA
GAGCCCAGAGCCATTGTCACACACCAAGGCAGT
mm3__chr6 101.43 Mbp_chr6_101431430 TGATGCTGTCCTGTGTGCTTACTAGAAAGACAAGTCTTTACTCTGACAGAGTTTATATTCTGGTGCTGGGGGCGCCCAAC
AATGGAGGGGAACATGGCTTGGGGGCGTCATCCCCAGCAAAGCCG
mm4__chrY 18.15 Mbp_chrY_18147967 ACAATGGCTCTTGCATGTGCAAAGCTGGCTTTGCAGGGGACGATGCTCCCCAAGCCATGTTTCCCTCCACCGTTGGGTTC
CCCGGACACCAGGGCGTGATGGTGGACATGGGCCAGAAGGACTC
mm4__chrY 17.76 Mbp_chrY_17758018 GAGTCCTTCTGGCCCATGTCCACCATCACGCCCTGGTGTCCGGGGAACCCAACGGTGGAGGGAAACATGGCTTGGGGAGC
ATCGTCCCCTGCAAAGCCAGCTTTGCACATGCAAGAGCCATTGT
mm4__chr16 5.51 Mbp_chr16_5508661 AGAGTGCCACACCCAAGATGTTGGGGGTGGGGGGACTGCACAGAGCGATTGTACAGAGTGCCACACCCAAGATGGTGGGG
AGGACTGCACAGAGCGATTGCACAGAGT
mm4__chr16 5.91 Mbp_chr16_5905904 CACTCCCCAAATGCCTCCTCCGGAAGCCTTCCTAATTTCCCTCTTTAGAATTCATCTCTTTTTCCCGACGATGGAGGATA
AGAAGGTTTTCAGCTCTCTTGTAGTGGGAATTGGCTGCCCAAAGCCCTACCTTAGG
mm4__chr16 50.06 Mbp_chr16_50059124 CGCCCACATAGGAGTCCTTCTGGCCCATCCATGCCCACCATCATGAGCTGGTGCCCGGGGCACCCGACAATGGAGGGGAA
CATGGTTCAGGGGGCATCGTCCCTAGCAAAGCCA
mm3__chr17 81.51 Mbp_chr17_81512247 CCATCCACTCACCTGGTGTCTGGGGCGCCCGACGATGGAAGGAAACACGGCTCGGGGAGCGTCGTCCCCAGCAAAACCAG
CTTTGCACATGCCGGAGCCATTGTCAATGACCAGCGCGGCGATCTCTTCTTCCAT
mm4__chr7 63.40 Mbp_chr7_63404892 AGGAAACTCATCAGCTTGGGTTTGATAAAAGGACGATTTAGAGGTATTCCCCTCCATCTTGGGGAGCGACCCTTGCAGCA
GAAAAATTGTGGTGAGAGTCTGGGGCAAATGGAGCCACTCCTTTTGCTGGAA
mm4__chr7 63.34 Mbp_chr7_63343195 TTCCAGCAAAAGGAGTGGCTCCATTTGCCCCAGACTCTCACCACAATTTTTCTGCTGCAAGGGTCGCTCCCCAAGATGGA
GGGGAATACCTCTAAATCGTCCTTTTATCAAACCCAAGCTGATGAGTTTCCT
mm2__chr1 229.43 Mbp_chr1_229433002 GCAGCCTGACCTGGTGTCGGGGGCGGCCCACGATGGACGGGAACACGGCCCTAGGGGCGTCATCCCCGGCGAAGCCGGCT
TTCACCAGGCCGGAGCCATTGTCGCACACGAGGGCGGTGGTCTCGTCTTCGTC
mm2__chr18 62.44 Mbp_chr18_62442092 CCTCGTCATCGACAATGGCTCTGGCATGTGCAGGGCTGGCTTCGCAGGCGACAGTGCTCTCCAGGCCGTCTTCCTCTCCA
TCATGGGGCACCCCAGGCACCAGGGTGTGATGGTGGGCATGGGTCAGAAGGACTC
mm4__chr1 237.93 Mbp_chr1_237926914 TGACAATGGCTCCAGCATGTGCAAAGCTGGCTTTGCTGGGGACGATGCCCCCCAAGCTGTGTTTCCCTCCATCATTGGGC
ACCCCCGGCACAAGAGCATGATGGTGGGCATGGGCCAGAAGGACTCCTACGTG
mm4__chr12 26.33 Mbp_chr12_26334148 CCAAGATCACAAAGATACACCTGACCCACGTTGAAGGGGAAGAGGGAGTGAGTTATGGTATAGAGAAAGGAGGATACAAT
TGTCCAAAGTGAAGATAATTATTACCAAAAAAACCAGAAAGCACAGTCATTAGAGCAGGG
mm4__chr10 85.62 Mbp_chr10_85617054 ATCCTGTCCTCTGCCAAAGCAAAGGAGCATATCTTTATTGAAGGAAATAGTTTAGTTCCCAGGATGAAGGGGAAGACAGT
TATAGCAGTTCCACATAAGGCTGGGACCTG
mm4__chr3 38.55 Mbp_chr3_38554277 TTCTCCGTCCAGCTGACTTGTATACCCACCCACGATGGAGAGGATGACAACCACGAAGTCGAAGATATTCCAGCTGTTGG
TGAAGTAGTAGTGGCGCAGGG
mm4__chr19 49.37 Mbp_chr19_49374719 CCTTGTTCTTCCTCCCCAGGTACCCAGGATGGAGGAGAAGGAGGCCCTGGTACCCATCCAGAAGGCCACGGACAGCTTCC
ACACAGAACTCCATCCCC
mm4__chr21 32.00 Mbp_chr21_31998305 AAGCAGGGACGATGTTAGGCTGGGGCCATGAGGCAAATTGGAATCCAGGAGACGTCCTGTCGCCAGCCCTCTGTCCCTCC
ATCCTAGGAAGACTGCCCGACCTCTGTGGGGAGAGTGACATGCTGCTGGT
mm4__chr12 126.88 Mbp_chr12_126884173 CAGGAGTCACAGAGTGGGGACCACCCAGCAAACGAAAGGCTGATTAGATGGGAGGCTGTCTTCCCCTCCAAAATGGGTCA
CATATCCCCCAAAGCAAGAGTTATTCTGG
mm4__chr19 40.40 Mbp_chr19_40398533 TGACAAGACAGAGGGCAAGGCTGGCCCACGATGGCGGGGAATGGGGCTCACGGCGCAGAGACCGGATCGCTGGGGCAGTC
CAGGGCCGGGGCCGGGCTAAGCACGCGTACCAGCTT
mm4__chr16 48.58 Mbp_chr16_48578495 GTTATCCCACACGTGCCTCAAACTTAACAGTTCTGAATCAGAGCTCACAGCTCTCTTCCACTCCATCCTTGGCCCAAGTT
GCCTTTCAGTGATGTGGGCACTGAGTCTTGTGA
mm4__chr10 129.19 Mbp_chr10_129192606 TGGTGACAGCACAGCAGTAGCACGGCTGGCCATTTCCTTCTTGTCCATCAGGCCGTCTCCACCTCCATCTGGGGGGCAGA
TTGGATCTTGCCAG
mm4__chr1 112.00 Mbp_chr1_112004369 TCTTTCTGCCCATAGCCTGGAAGGGCACAGGCCCAGGATGGAGGGCAAGAAGGATAAGGGATTGCTGCCCTGGGAAGCCT
ACATGTGTTTCCTGATCCAGAGGTCTTCCTCAGAGGCAAACA
mm4__chr17 79.10 Mbp_chr17_79104967 ACTGTTCAGGTCGTTTGGGCCAGCTGGGCCATGACGCTCTAGCTGCCTTCTTCCCTCCCATTGTTGGCCATAGTCCCCAT
GGGGCAGCCCTGTCTCTCTCACTG
mm3__chr5 147.47 Mbp_chr5_147466725 AAGCCAACCCCAAAGTGTGAGGCTGCAATCCCACTTTGTATTTTGCAGGCTTCCCCTCCATCTTCAGGAAGGGACTTTGG
CTGTGTTGT
mm4__chr1 61.74 Mbp_chr1_61743012 TATTTTGCGCCCAGATCGGTGCCCCTGTTTCGCTCCCAGAGCTGGGCGGAGTCTTTCCCTCCATCGGTGGCCGTGGAAGG
AGGGAACGGGGTGAGGGGACAAAGAGGAGGG
mm4__chr11 2.41 Mbp_chr11_2412146 ACCTGAGTGGCTTCATCTGGAAGCCTGCCCACAAGCCGCCCTGAGGCCACACGGCCTCTGGGCCCCACAGACCTGCAGGT
GACACCCACGATGAACAGGAAGATGGCCACCATGTCACACTTGTTCCAGTTGTCCCC
mm4__chr13 112.88 Mbp_chr13_112883403 TGCCTTTCAGGAAAGCACCACCAACGCTGGAGGAGGAGCCGGCCCTCACGCCCGCCCCGCGCCACGCTGTGGAACGGGGC
TCCGGCAAGTGAAACCCAGAGGGTGTTTCCGAGGTG
mm3__chr1 223.86 Mbp_chr1_223864744 TGACCCATGCTCACCATCACGCCCTGGTGCCTGGGGCGCCCCACCATGGAAGGGAAGATGGCCCAGGGGTCATCATCCCC
TGTGAAGCCAGCCTTGCACATGCCGGAGCTGTTGTCAATGACGAGCG
mm4__chr11 111.28 Mbp_chr11_111284161 GGTCACGTCAGGTTACTACGGTGTCAGAAGATCTTTCTTATCTGACTCAGACTTCCACAACAGTAAACAGTTTTCAAATG
ACGTCTACACCTCCAGCGTGGGGAAGCCGTTTCCCTGTGAGTCCTC
mm2__chr3 12.07 Mbp_chr3_12070308 TCATTGACAGTGTCTCCGGCATGTACAAAGCTGGATTTGCTGGGGACGACGCCCCCTGAGCCGTGTTCCCCTCCATCCTT
AGGCGCCCCTGGCACCAGGGCACGATGGTGGGCATGGGTCAGAAGGACTC
mm3__chr12 89.12 Mbp_chr12_89123197 ACCAGATGAATATGTTAAATGCACCTTACAATATTTGAGATCTTCAAAAGAAAAAGATGGAGCTCACGTTGGAGGGGAAG
AAGGAAGAAAGAGGAGGAATGGAAAAGGGAGTAGAATGCTTTTCTTGGGTGTACATGC
mm4__chr16 49.52 Mbp_chr16_49524459 AGTGTGGCAGTGACAGATGGGTGGGGGCTCCTGCGATGAGGGGGAAGATGGCATCAGGGTGCTCAGGGGCTCTGGGGAGC
CCCAGGGGTGTGTGTTTGGGGGATTTGCCCCAGAGTGGACA
mm4__chr20 50.24 Mbp_chr20_50243254 GGCACAAAGGCCCATTGTTCGGGCCCACGTTGGCGGGGAAGCGGGCGGGGAGCAGCCGCCTTTCACCCAACCCGCTTGTG
GTCGGCGGGGACTGAGGAGCGATTCAC
mm4__chr2 27.37 Mbp_chr2_27373994 AGGATGTCCTGGAGGTGAGGCGCTTGTTCAGCACTGCCCCTTCTTCCCCTACATCCTGGGGTCCTGGGCTTGGAGAATGA
TTGGAACCAGCCAGTATGATGAGCTGTACTTCTATCCCTATCCCCAGGCTGTAGCTG
mm4__chr18 58.26 Mbp_chr18_58255782 TTCATGGCATTCCACACCCTACCGCGGAGAAGCAGCCCGCACCCCTTCGCCCAGAACGGAGGGGAGGACGGCGGCGGAGG
CCTGCAGGGAGGCGTGGGTGCGCTTAAGCGGAGCTCGTCCAT
mm4__chr9 93.95 Mbp_chr9_93947410 TCCCCTGTGAATTCCTTGGCCTAGCTGAAGGACTCCCCTCTGTGGTCTTCCCCTCCAGTGTTGGTGCCTGAGAGGCCAGG
GGCCACCACTGTTCACTGGAATGGCTAAATAACCTCGGCAGTACACCCATGGGAGTGG
mm4__chr5 57.48 Mbp_chr5_57482597 ATCTCCCACGTAGCAGTCCTTCTGGCCCATGCCTACCATAACGCCCTGGTGTCGAGGACGCCCTATCATGGAGGGGAACA
CAGCCCGGGGGGCATCGTCACCACCAAAGCCTGCCTTGCACATCCCTGACCCAT
mm4__chr8 84.95 Mbp_chr8_84948672 CTCTGGTATGTGCAAGGCCAGAATCCTTGTCAAAGATGCCCCCAGGCCTTCTTCCTTTCCATAGTGGGATGCCCATGGGA
CCAGAGTTTGATGATGGGCATAGGGCAGAAGGACTG
mm4__chr1 202.87 Mbp_chr1_202873375 TGCTGGGTATAGTTCCCCGAGCCATGTTCCCCTCCATCACTGGGTATCCCTGGCACCAGGGCATGATGGTGGGCATAGGC
CAGAAGGACTCC
mm4__chr19 3.99 Mbp_chr19_3985433 GAACCGAAAGAAGCGAGTCGCGCCGAGGATGGCGGCGACGACGGCGGAAGAGAACGCTGACGTCAACACTCAGCT
mm3__chr2 131.63 Mbp_chr2_131626222 GTGCTCGTCATTGACAACGGCTCTGGCATGTGCAAGGCCGGCTTTGCGGGCGACGATGCCCCCCAGGCTGTCTTCCCTTC
TATCGTGGGGCGCCCCAGGCACCAGGGCATGATGGAGGGCATGCATCAGAAGGAGT
mm4__chr20 57.73 Mbp_chr20_57734279 GCTGCCCCATCTCAAGGAATAAGCCTTCTTCCCCTTCAGCTTTGGCAGATGGCCCTGGCTTTTGCACACCTGGTCTC
mm4__chr11 112.53 Mbp_chr11_112529298 TGAGGAGGGTAGCACCAGAATTTTTCCAAGGCAACTGCAGCCTCATGACCTGTGATGGAGGGAAAGAAGGCTGGTGATGG
TAGCCAGCCAGCCTGCTGAGTGGCCCGTGATTGTACA
mm4__chr5 5.73 Mbp_chr5_5725799 ATCTGGGGCATGGAAATGGGCTGCAGCTCCAGGACCTTCTTGAATCCCACCATGTAGGGGAACACTGCCCTGGAGGTAGG
GATCTTGAAATTCTGCCTTTTCAAGAGCAGCA
mm4__chr5 98.34 Mbp_chr5_98338826 GCTCCTCATCAACAAGGGCTCTGGCATGTGCAAGGCCGGCTTCACAGGTGACAATGCCCCCGGGCCATCTTCCCCTCTGT
CATGGGTGCCAGGGAGTGATGGTGGGCATGGGTCAAAA
mm4__chr16 2.52 Mbp_chr16_2523272 CTAGTTCTGTCCAGAGCCCCCGCCGTTTTCCCCTCCAGCACAGGATTCATTTCAGGATCATCACTTGCGTTATATGCCCA
AGGGGTCCCTCTCTTGGTGTGTG
mm4__chr19 54.14 Mbp_chr19_54141816 TGACTACTCCAGGGTTCCCTGCCCTGCACACGTAGAGTGGGAGCGGCCCGTGGATCCCAGGGAACTGTGCTTTTCATTGT
AGGCCCCCTCCCTGGAGGGGAAGAGGGCAATCTCCGCTGGTATCTCAGAAGTCT
mm4__chr19 33.41 Mbp_chr19_33406847 CCATGTGAGGCCAAGGTGGCTGCTGAGGTCCTGGGTGAGCGGGCACCCCATGGCCTGCAGGGTCGAGGTCTCGCCATTAT
CTAGCCAGGGAAGGAGGGGAAGAGGGCACTGGTGGCTGCTGAGTGTGAGGGCTG
mm4__chr16 87.44 Mbp_chr16_87436982 TCTGCACGCAGACACTCATTACAACCCACTTCCAGTCTGCCCCCAGTGTGGCCGCCTTCCCCTCCCTCCCTGGTCAGCCC
CACTGCCTTCTGTGGAGCGTAAG
mm4__chr11 2.27 Mbp_chr11_2269483 GATCCGCCAGCACTAGAGACCTCAACGGCCCGACGGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTCGTCGGCGGCTGAG
CCGCGAGGGGAAGTTTTGCAATCCCGGACAAACAAACGCCGGTCTTG
mm4__chr1 46.13 Mbp_chr1_46133074 GCGTCTTTTGTCTGCCTTGCTCCTGCGAAGGAGCGGGAGACGGCTGCGCGCCCCAAGCGGGCCAATCAGGAGACAGCCGG
CGCCGGCCGGTAAGGCGGCGATTGGTCAGAGCCCAGTTACTAAGCGGGACAAAGGCAGA
mm4__chr1 215.08 Mbp_chr1_215081843 TCGCCTATTACGACGACAGCAACGTCTGATCTCCTGGCTTTCTTCCCCTCCTTGGTGGGTTCCGTTCCGTCCCCCTGTCG
CTGCGAGTTTGGCTGTCCCTACG
mm4__chr1 1.98 Mbp_chr1_1977389 CCCTCTGCATATCCGTCCACAGGGAAGCAGCCTTCCCCTCCCTCGAAGGCAGGCTCTGCTGCCTGGGGGCCCCGTAACAC
CTCCCTGAGGGGCTGCCAGAGCTCCTCATGACCCCCTTGCCATCCTCTCTGGCCCAGATG
mm4__chr1 1.60 Mbp_chr1_1600886 CAGGGCTGGACAAAGGGAATCCAAAAAGCCCACCCGGTGCACTGAGTAGGTGGCACCGACGGGGGAGGGGAAGGCGGGAC
ACACCCCAAGACCCCTGCCCCTCATTCCCGTCCCTGCATAGATCAC

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.