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Contents:

PCR primers for off-targets of CCCGGGTCGCGGGGACGGCG CGG

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.

If a primer was not found, the reason is usually that the region around the off-target is too repetitive. To avoid unspecific primers, all repeats are masked for the primer design (not for off-target search). If you think that we should change the parameters here or should use different primer3 settings, please let us know.

Maximum amplicon length:     Primer Tm:

NamePrimer Sequence Tm CFD Score
mm0_exon_Pop5_chr5_115238085_F TCGTCGGCAGCGTCTGCGGTTCAAGCACAGGTAA 60.1 1.00
mm0_exon_Pop5_chr5_115238085_R GTCTCGTGGGCTCGGCTCGCACAGAAGGTACCTGC 60.7 1.00
mm4_exon_Agpat2_chr2_26603918_F TCGTCGGCAGCGTCCTACAAGCGGCGCACATC 59.2 0.64
mm4_exon_Agpat2_chr2_26603918_R GTCTCGTGGGCTCGGCCTCGCGTTCTTTCCTTCCT 60.0 0.64
mm3_exon_Fibcd1_chr2_31845983_F Primer3: not found at this Tm N.d. 0.60
mm3_exon_Fibcd1_chr2_31845983_R Primer3: not found at this Tm N.d. 0.60
mm4_intergenic_Banp|Gm22_chr8_122172637_F TCGTCGGCAGCGTCCCATTCACCCTACAGCCTGG 60.1 0.60
mm4_intergenic_Banp|Gm22_chr8_122172637_R GTCTCGTGGGCTCGGCCAGCTGCAGAACCTTCAGA 59.9 0.60
mm3_intron_Pmp22_chr11_63132535_F TCGTCGGCAGCGTCGGCTGAAGGCTCTTCACCC 60.3 0.58
mm3_intron_Pmp22_chr11_63132535_R GTCTCGTGGGCTCGGCTCGGCCTAGATGCTGGC 59.9 0.58
mm4_intron_Aff4_chr11_53351182_F TCGTCGGCAGCGTCGGGAAATGGTTCGGGCCTAG 60.4 0.49
mm4_intron_Aff4_chr11_53351182_R GTCTCGTGGGCTCGGGGTCTCCGCCAGCGTCAG 62.8 0.49
mm4_intergenic_Dpy19l1|Dpy19l2_chr9_24503221_F TCGTCGGCAGCGTCCTCACTCTGGCCGGCGAG 62.5 0.47
mm4_intergenic_Dpy19l1|Dpy19l2_chr9_24503221_R GTCTCGTGGGCTCGGCCCTGGCAACCCGAGAGC 62.7 0.47
mm4_exon_Kcnb1_chr2_167189986_F Primer3: not found at this Tm N.d. 0.40
mm4_exon_Kcnb1_chr2_167189986_R Primer3: not found at this Tm N.d. 0.40
mm4_intergenic_Gm9727|Btbd8_chr5_107479359_F Primer3: not found at this Tm N.d. 0.40
mm4_intergenic_Gm9727|Btbd8_chr5_107479359_R Primer3: not found at this Tm N.d. 0.40
mm4_intergenic_Kalrn|Ropn1_chr16_34573926_F TCGTCGGCAGCGTCCTAGGCCGGCTTCCTGAG 59.1 0.35
mm4_intergenic_Kalrn|Ropn1_chr16_34573926_R GTCTCGTGGGCTCGGGACCCGGAGCTTGCGGAG 62.8 0.35
mm4_intron_Gata4_chr14_63255791_F Primer3: not found at this Tm N.d. 0.35
mm4_intron_Gata4_chr14_63255791_R Primer3: not found at this Tm N.d. 0.35
mm4_exon_C1qc_chr4_136890572_F Primer3: not found at this Tm N.d. 0.34
mm4_exon_C1qc_chr4_136890572_R Primer3: not found at this Tm N.d. 0.34
mm4_intergenic_Gm11746|Slc24a3_chr2_145167692_F Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Gm11746|Slc24a3_chr2_145167692_R Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Cmpk2|Gm9866_chr12_26486850_F Primer3: not found at this Tm N.d. 0.32
mm4_intergenic_Cmpk2|Gm9866_chr12_26486850_R Primer3: not found at this Tm N.d. 0.32
mm3_intergenic_Chrdl2|Pold3_chr7_100042276_F TCGTCGGCAGCGTCACGGGCTTTGATTCGTGACT 59.9 0.30
mm3_intergenic_Chrdl2|Pold3_chr7_100042276_R GTCTCGTGGGCTCGGAACAAACACGTCCCACAAGC 59.5 0.30
mm4_exon_Slc7a5_chr8_121907491_F Primer3: not found at this Tm N.d. 0.29
mm4_exon_Slc7a5_chr8_121907491_R Primer3: not found at this Tm N.d. 0.29
mm4_exon_Fbxl16_chr17_25819903_F Primer3: not found at this Tm N.d. 0.27
mm4_exon_Fbxl16_chr17_25819903_R Primer3: not found at this Tm N.d. 0.27
mm4_intergenic_Gm12904|1700121C08Rik_chr4_123842070_F Primer3: not found at this Tm N.d. 0.27
mm4_intergenic_Gm12904|1700121C08Rik_chr4_123842070_R Primer3: not found at this Tm N.d. 0.27
mm4_exon_Sept3_chr15_82275300_F Primer3: not found at this Tm N.d. 0.26
mm4_exon_Sept3_chr15_82275300_R Primer3: not found at this Tm N.d. 0.26
mm4_intron_Zfp398_chr6_47835952_F TCGTCGGCAGCGTCGGTAAGGGCGCTGATGGAC 60.5 0.23
mm4_intron_Zfp398_chr6_47835952_R GTCTCGTGGGCTCGGCAAACGACAGCCGCAGAC 59.4 0.23
mm4_intergenic_Gzmk|BC067074_chr13_113293228_F TCGTCGGCAGCGTCCGCGCAGGTAGAGCTCGG 62.9 0.23
mm4_intergenic_Gzmk|BC067074_chr13_113293228_R GTCTCGTGGGCTCGGCCCCAGAAAGGCGGTGAG 60.0 0.23
mm4_intergenic_Gm12249|Larp1_chr11_58008859_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Gm12249|Larp1_chr11_58008859_R Primer3: not found at this Tm N.d. 0.22
mm3_intron_Rnf130_chr11_50025677_F Primer3: not found at this Tm N.d. 0.22
mm3_intron_Rnf130_chr11_50025677_R Primer3: not found at this Tm N.d. 0.22
mm4_exon_Tm9sf3_chr19_41263990_F Primer3: not found at this Tm N.d. 0.22
mm4_exon_Tm9sf3_chr19_41263990_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Gm24720|Tshz1_chr18_83936519_F Primer3: not found at this Tm N.d. 0.21
mm4_intergenic_Gm24720|Tshz1_chr18_83936519_R Primer3: not found at this Tm N.d. 0.21
mm4_exon_Rbm15_chr3_107332581_F TCGTCGGCAGCGTCCCAGCTCGCTGATCTTCAGG 60.5 0.21
mm4_exon_Rbm15_chr3_107332581_R GTCTCGTGGGCTCGGGAGTCACGTTCCTCCGGG 59.4 0.21
mm4_intron_6330403A02Rik_chr1_180483062_F TCGTCGGCAGCGTCGACACATCCACGGGTCTCAG 60.1 0.19
mm4_intron_6330403A02Rik_chr1_180483062_R GTCTCGTGGGCTCGGGTGCCGGGATTGGGTAAGAA 60.0 0.19
mm4_intron_Mdm1_chr10_118141981_F Primer3: not found at this Tm N.d. 0.19
mm4_intron_Mdm1_chr10_118141981_R Primer3: not found at this Tm N.d. 0.19
mm3_intron_Grlf1_chr7_16614282_F Primer3: not found at this Tm N.d. 0.18
mm3_intron_Grlf1_chr7_16614282_R Primer3: not found at this Tm N.d. 0.18
mm4_exon_Nudt9_chr5_104047125_F TCGTCGGCAGCGTCTCAGTGACTGTCCGGTCCTC 60.8 0.18
mm4_exon_Nudt9_chr5_104047125_R GTCTCGTGGGCTCGGAAAAGGAAGGAGGGGATCGC 59.7 0.18
mm4_exon_Tbc1d2b_chr9_90270686_F Primer3: not found at this Tm N.d. 0.18
mm4_exon_Tbc1d2b_chr9_90270686_R Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Gm13780|Phf21a_chr2_92183681_F TCGTCGGCAGCGTCCCCAGCTCCAGTCCCTCT 59.9 0.17
mm4_intergenic_Gm13780|Phf21a_chr2_92183681_R GTCTCGTGGGCTCGGCAGGCCCAGACACACTCG 60.0 0.17
mm3_exon_Itga5_chr15_103366674_F Primer3: not found at this Tm N.d. 0.17
mm3_exon_Itga5_chr15_103366674_R Primer3: not found at this Tm N.d. 0.17
mm4_exon_Fbxo30_chr10_11281570_F Primer3: not found at this Tm N.d. 0.16
mm4_exon_Fbxo30_chr10_11281570_R Primer3: not found at this Tm N.d. 0.16
mm3_intergenic_Elmod1|Tnfaip8l3_chr9_53975358_F TCGTCGGCAGCGTCGTTTCCCTGGGAGCAAGACG 60.9 0.16
mm3_intergenic_Elmod1|Tnfaip8l3_chr9_53975358_R GTCTCGTGGGCTCGGGGCCAAGTGCGCAGTTTTAA 59.9 0.16
mm4_exon_Gclc_chr9_77754636_F Primer3: not found at this Tm N.d. 0.16
mm4_exon_Gclc_chr9_77754636_R Primer3: not found at this Tm N.d. 0.16
mm4_intron_Ahi1_chr10_20952794_F TCGTCGGCAGCGTCCTCTCTGTGAGTATCCCGCG 59.6 0.16
mm4_intron_Ahi1_chr10_20952794_R GTCTCGTGGGCTCGGGAGCATCCCACAGTCTCCG 59.8 0.16
mm4_exon_Atp8b1_chr18_64660909_F TCGTCGGCAGCGTCGACTCCTCAGGCTCTCGTGG 61.6 0.15
mm4_exon_Atp8b1_chr18_64660909_R GTCTCGTGGGCTCGGCCCTGGAGCAGAGAAACCAG 60.0 0.15
mm4_intron_Rnf219_chr14_104522373_F Primer3: not found at this Tm N.d. 0.15
mm4_intron_Rnf219_chr14_104522373_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Zfand4|March8_chr6_116337931_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Zfand4|March8_chr6_116337931_R Primer3: not found at this Tm N.d. 0.15
mm4_exon_Sbk1_chr7_126273071_F Primer3: not found at this Tm N.d. 0.14
mm4_exon_Sbk1_chr7_126273071_R Primer3: not found at this Tm N.d. 0.14
mm3_exon_Gm26776/Ccdc3_chr2_5137874_F TCGTCGGCAGCGTCTAGCCGTGCACCCAGCTC 62.0 0.14
mm3_exon_Gm26776/Ccdc3_chr2_5137874_R GTCTCGTGGGCTCGGCTACGCTCTAGCCCGGGC 61.9 0.14
mm4_intergenic_Gm9755|Nat3_chr8_67523119_F Primer3: not found at this Tm N.d. 0.14
mm4_intergenic_Gm9755|Nat3_chr8_67523119_R Primer3: not found at this Tm N.d. 0.14
mm4_exon_B4galt3_chr1_171270381_F TCGTCGGCAGCGTCCCAAGATGGCTGCAGCGATA 60.5 0.13
mm4_exon_B4galt3_chr1_171270381_R GTCTCGTGGGCTCGGACTCTCTCTCTCCGTGGTCC 60.0 0.13
mm4_intron_Slc2a1_chr4_119109249_F Primer3: not found at this Tm N.d. 0.13
mm4_intron_Slc2a1_chr4_119109249_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_C230037L18Rik|Shank3_chr15_89499579_F Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_C230037L18Rik|Shank3_chr15_89499579_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Ppm1h|Mirlet7i_chr10_122985563_F TCGTCGGCAGCGTCCGGGGAGCTAACGGTTTCC 60.4 0.13
mm4_intergenic_Ppm1h|Mirlet7i_chr10_122985563_R GTCTCGTGGGCTCGGGCTACTGCCTTGCTAGTGCT 60.1 0.13
mm4_intergenic_Gm26843/Gnao1|Gnao1_chr8_93810429_F Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Gm26843/Gnao1|Gnao1_chr8_93810429_R Primer3: not found at this Tm N.d. 0.12
mm4_exon_Cpeb2/Gm7854_chr5_43234703_F Primer3: not found at this Tm N.d. 0.12
mm4_exon_Cpeb2/Gm7854_chr5_43234703_R Primer3: not found at this Tm N.d. 0.12
mm4_intron_Usp9x_chrX_13072730_F Primer3: not found at this Tm N.d. 0.11
mm4_intron_Usp9x_chrX_13072730_R Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Camk2n1|Vwa5b1_chr4_138462806_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Camk2n1|Vwa5b1_chr4_138462806_R Primer3: not found at this Tm N.d. 0.11
mm4_exon_Ralgds_chr2_28533476_F Primer3: not found at this Tm N.d. 0.11
mm4_exon_Ralgds_chr2_28533476_R Primer3: not found at this Tm N.d. 0.11
mm4_exon_Igf2bp3_chr6_49214416_F TCGTCGGCAGCGTCCCGGGATCTTAGCGTCCTTG 60.2 0.10
mm4_exon_Igf2bp3_chr6_49214416_R GTCTCGTGGGCTCGGTCGGGTTTTAAAACGACCCCT 59.8 0.10
mm4_intron_Snx6_chr12_54795580_F TCGTCGGCAGCGTCAAGAGAGACATTAGCGCCCA 58.8 0.10
mm4_intron_Snx6_chr12_54795580_R GTCTCGTGGGCTCGGGGACTCGAAGCAGCCATGAT 60.1 0.10
mm4_intron_Hdac2_chr10_36974825_F Primer3: not found at this Tm N.d. 0.10
mm4_intron_Hdac2_chr10_36974825_R Primer3: not found at this Tm N.d. 0.10
mm3_intron_Gm15608_chr10_82178505_F TCGTCGGCAGCGTCCGGAATCCGCACCATGGT 60.1 0.10
mm3_intron_Gm15608_chr10_82178505_R GTCTCGTGGGCTCGGGAGCGTGAGTTCCAGGGAC 60.0 0.10
mm4_intron_Eif5a2_chr3_28781426_F Primer3: not found at this Tm N.d. 0.09
mm4_intron_Eif5a2_chr3_28781426_R Primer3: not found at this Tm N.d. 0.09
mm3_intergenic_Atp11c|Gm7073_chrX_60404065_F Primer3: not found at this Tm N.d. 0.09
mm3_intergenic_Atp11c|Gm7073_chrX_60404065_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Usp2_chr9_44084846_F TCGTCGGCAGCGTCAGGGTCATCGGCTTTGGG 59.3 0.09
mm4_intron_Usp2_chr9_44084846_R GTCTCGTGGGCTCGGCAGCTTTGCGACGTCAGC 59.8 0.09
mm4_exon_Ppp2r3d_chr9_124424000_F Primer3: not found at this Tm N.d. 0.09
mm4_exon_Ppp2r3d_chr9_124424000_R Primer3: not found at this Tm N.d. 0.09
mm4_exon_Cacna1b_chr2_24762962_F Primer3: not found at this Tm N.d. 0.08
mm4_exon_Cacna1b_chr2_24762962_R Primer3: not found at this Tm N.d. 0.08
mm4_exon_Yy1_chr12_108793966_F Primer3: not found at this Tm N.d. 0.08
mm4_exon_Yy1_chr12_108793966_R Primer3: not found at this Tm N.d. 0.08
mm4_intron_Il17d_chr14_57525157_F Primer3: not found at this Tm N.d. 0.08
mm4_intron_Il17d_chr14_57525157_R Primer3: not found at this Tm N.d. 0.08
mm4_intron_Cd63_chr10_128910154_F Primer3: not found at this Tm N.d. 0.07
mm4_intron_Cd63_chr10_128910154_R Primer3: not found at this Tm N.d. 0.07
mm4_intron_Sar1a_chr10_61680479_F Primer3: not found at this Tm N.d. 0.07
mm4_intron_Sar1a_chr10_61680479_R Primer3: not found at this Tm N.d. 0.07
mm4_exon_Dennd1b_chr1_138963686_F Primer3: not found at this Tm N.d. 0.06
mm4_exon_Dennd1b_chr1_138963686_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm12341|Abr_chr11_76562016_F TCGTCGGCAGCGTCAAAGCTCTGGGCAGTTGAGG 60.2 0.06
mm4_intergenic_Gm12341|Abr_chr11_76562016_R GTCTCGTGGGCTCGGTCCTGGGGTCCTGTTCTGTC 60.8 0.06
mm4_intergenic_Ppp3r1|Wdr92/Ppp3r1_chr11_17159798_F TCGTCGGCAGCGTCCTCCCCGAGATTCGCCGC 62.9 0.06
mm4_intergenic_Ppp3r1|Wdr92/Ppp3r1_chr11_17159798_R GTCTCGTGGGCTCGGAAAAGGCCGAACGAGCCAG 60.9 0.06
mm4_intergenic_Gm10797|Zfp365_chr10_67777114_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm10797|Zfp365_chr10_67777114_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gab1|Gm27048/Gm4891_chr8_80880608_F TCGTCGGCAGCGTCGGGATCTGTAGTCAGCAGGC 59.8 0.06
mm4_intergenic_Gab1|Gm27048/Gm4891_chr8_80880608_R GTCTCGTGGGCTCGGCACTCCCGCTCTCACACTG 60.0 0.06
mm4_exon_Tbc1d4_chr14_101608150_F TCGTCGGCAGCGTCTGCTCGAAGATGAACACCCC 60.0 0.06
mm4_exon_Tbc1d4_chr14_101608150_R GTCTCGTGGGCTCGGGAGGTGATCCTGGTGCTCAG 59.8 0.06
mm4_intergenic_Gm16195|Srsf3_chr17_29032421_F TCGTCGGCAGCGTCTAGGCGGAAGTGGAGGAGAG 60.3 0.06
mm4_intergenic_Gm16195|Srsf3_chr17_29032421_R GTCTCGTGGGCTCGGGTTCCGTCAGGTGGGTGG 59.9 0.06
mm4_exon_Zfp664_chr5_124863026_F TCGTCGGCAGCGTCACAACCCATCCTTCAACCGG 60.2 0.06
mm4_exon_Zfp664_chr5_124863026_R GTCTCGTGGGCTCGGGCCTCTTCTGCCTCATCCC 59.8 0.06
mm4_intergenic_Kcnj8|Abcc9_chr6_142571399_F TCGTCGGCAGCGTCAGGTGCGATCCATAGTGCAC 60.1 0.06
mm4_intergenic_Kcnj8|Abcc9_chr6_142571399_R GTCTCGTGGGCTCGGCACCTCTATCCCCAGCTCCT 60.1 0.06
mm4_intron_Nr5a1_chr2_38711805_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Nr5a1_chr2_38711805_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Slc35f1|Gm25664_chr10_52691567_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Slc35f1|Gm25664_chr10_52691567_R Primer3: not found at this Tm N.d. 0.05
mm4_exon_Shank3_chr15_89499735_F Primer3: not found at this Tm N.d. 0.05
mm4_exon_Shank3_chr15_89499735_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Grin2b/Gm26653|Grin2b_chr6_136173328_F TCGTCGGCAGCGTCCAGCAGGAGAGGAAGAGCAG 59.8 0.05
mm4_intergenic_Grin2b/Gm26653|Grin2b_chr6_136173328_R GTCTCGTGGGCTCGGCACGTTCCTGGGTGTGAGAC 60.6 0.05
mm4_exon_Brd3_chr2_27475364_F Primer3: not found at this Tm N.d. 0.05
mm4_exon_Brd3_chr2_27475364_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Gm13012|Ece1_chr4_137897857_F TCGTCGGCAGCGTCGCCCTCAGTTTTCCCATCCC 60.6 0.05
mm4_intergenic_Gm13012|Ece1_chr4_137897857_R GTCTCGTGGGCTCGGTGTCCCCATCCACAAATGCA 59.8 0.05
mm4_intergenic_Chd6|Gm14243_chr2_161109341_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Chd6|Gm14243_chr2_161109341_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Gm5229|Crim1_chr17_78200142_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Gm5229|Crim1_chr17_78200142_R Primer3: not found at this Tm N.d. 0.04
mm4_exon_Scarf2_chr16_17797394_F Primer3: not found at this Tm N.d. 0.04
mm4_exon_Scarf2_chr16_17797394_R Primer3: not found at this Tm N.d. 0.04
mm4_exon_Nf1_chr11_79339980_F Primer3: not found at this Tm N.d. 0.04
mm4_exon_Nf1_chr11_79339980_R Primer3: not found at this Tm N.d. 0.04
mm4_exon_Arhgef16_chr4_154299870_F TCGTCGGCAGCGTCGTCTTCCCCTTGGCTGACTC 60.0 0.04
mm4_exon_Arhgef16_chr4_154299870_R GTCTCGTGGGCTCGGAGGAAGTAGAGAGGGCGGG 60.0 0.04
mm4_intron_Rala_chr13_17943812_F TCGTCGGCAGCGTCCTTGAAAGCGAGGGAGGTCG 60.7 0.03
mm4_intron_Rala_chr13_17943812_R GTCTCGTGGGCTCGGAAGGTCACCTGTATAGCGCC 59.5 0.03
mm3_intron_Sctr_chr1_120007340_F TCGTCGGCAGCGTCCTGCTGCTCATCAATGCAGC 60.2 0.03
mm3_intron_Sctr_chr1_120007340_R GTCTCGTGGGCTCGGGAAGAGAGATCGGCTGCTCC 59.9 0.03
mm4_intergenic_Ccdc105|Slc1a6_chr10_78780364_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Ccdc105|Slc1a6_chr10_78780364_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Gm8770_chrX_78785863_F Primer3: not found at this Tm N.d. 0.03
mm4_exon_Gm8770_chrX_78785863_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Ephb2_chr4_136835893_F Primer3: not found at this Tm N.d. 0.03
mm4_exon_Ephb2_chr4_136835893_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Tm2d3|Gm7551_chr7_65702966_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Tm2d3|Gm7551_chr7_65702966_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm25266|Cradd_chr10_95136497_F TCGTCGGCAGCGTCCCATGAAGTGGTGGTGTTTTCC 59.9 0.03
mm4_intergenic_Gm25266|Cradd_chr10_95136497_R GTCTCGTGGGCTCGGTTTCTGCTCCCACAGGCATC 60.3 0.03
mm4_intron_Fryl_chr5_73255904_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Fryl_chr5_73255904_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Homer3_chr8_70283048_F Primer3: not found at this Tm N.d. 0.03
mm4_exon_Homer3_chr8_70283048_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Usp15|Usp15/AC158802.1_chr10_123196796_F TCGTCGGCAGCGTCGACCTGTCCGCACCTGTC 60.0 0.03
mm4_intergenic_Usp15|Usp15/AC158802.1_chr10_123196796_R GTCTCGTGGGCTCGGTCAAAACCTCGCTCCGGAAA 59.8 0.03
mm4_intron_Cep135_chr5_76588958_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Cep135_chr5_76588958_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Kmt2d|Rhebl1_chr15_98871542_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Kmt2d|Rhebl1_chr15_98871542_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Slc45a4_chr15_73645506_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Slc45a4_chr15_73645506_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Gm10657/Cd276_chr9_58555036_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Gm10657/Cd276_chr9_58555036_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gria4|Gm24896_chr9_4977682_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gria4|Gm24896_chr9_4977682_R Primer3: not found at this Tm N.d. 0.02
mm2_exon_Peli1_chr11_21091606_F TCGTCGGCAGCGTCCTGGTGCCTGCGTTCAAC 59.3 0.02
mm2_exon_Peli1_chr11_21091606_R GTCTCGTGGGCTCGGCTCACCGGACGACTGCTG 60.1 0.02
mm4_intergenic_Suv39h1|Suv39h1/Gm14820_chrX_8074373_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Suv39h1|Suv39h1/Gm14820_chrX_8074373_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Drd4_chr7_141294210_F TCGTCGGCAGCGTCTTTACTGGGCCACTTTCCGC 60.8 0.02
mm4_exon_Drd4_chr7_141294210_R GTCTCGTGGGCTCGGGCAGTCTGGGAAGAAGGGAC 60.0 0.02
mm4_exon_Fam211a_chr11_62648020_F TCGTCGGCAGCGTCGACAGGTCAGCCTTCCTCAC 60.0 0.02
mm4_exon_Fam211a_chr11_62648020_R GTCTCGTGGGCTCGGGGGTGACAGGAGAGCGCC 62.4 0.02
mm4_intergenic_Bend6|Dst_chr1_33908183_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Bend6|Dst_chr1_33908183_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Prkcb_chr7_122289278_F TCGTCGGCAGCGTCCTAGTCCCGCGCCTTTCC 60.2 0.02
mm4_exon_Prkcb_chr7_122289278_R GTCTCGTGGGCTCGGCCGGGTCAGCCATCTTGC 60.8 0.02
mm4_intergenic_Evx2|Hoxd13_chr2_74667870_F TCGTCGGCAGCGTCGGGTGGCTGCAAAGTCAACT 61.1 0.02
mm4_intergenic_Evx2|Hoxd13_chr2_74667870_R GTCTCGTGGGCTCGGCACCCCTCTTCAGCCCTTTG 60.6 0.02
mm3_intron_Rab31_chr17_65729985_F Primer3: not found at this Tm N.d. 0.02
mm3_intron_Rab31_chr17_65729985_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_N4bp2l1|N4bp2l2_chr5_150594564_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_N4bp2l1|N4bp2l2_chr5_150594564_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Eif4h|Limk1_chr5_134639368_F TCGTCGGCAGCGTCGTCTCCGCTCCTTGAGGAAC 60.1 0.01
mm4_intergenic_Eif4h|Limk1_chr5_134639368_R GTCTCGTGGGCTCGGGACTTGCGATCAGGCAGCG 61.8 0.01
mm4_intergenic_Fbxo11|Gm4832_chr17_88065366_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Fbxo11|Gm4832_chr17_88065366_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_4930515G01Rik/Ankrd13a_chr5_114774982_F TCGTCGGCAGCGTCTCGGTTCGCGAGCTGTCA 61.6 0.01
mm4_exon_4930515G01Rik/Ankrd13a_chr5_114774982_R GTCTCGTGGGCTCGGCAAGGGCCTCCGAGTCCG 62.1 0.01
mm4_intron_Mgat5_chr1_127304727_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Mgat5_chr1_127304727_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_BC052040|Meis2_chr2_115859176_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_BC052040|Meis2_chr2_115859176_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Actg1/mmu-mir-6935_chr11_120347905_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Actg1/mmu-mir-6935_chr11_120347905_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Zfp644/RP24-421H18.2_chr5_106696263_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Zfp644/RP24-421H18.2_chr5_106696263_R Primer3: not found at this Tm N.d. 0.00

Off-target amplicon sequences with primers

These only list off-targets that have primers in the table above. Primers underlined, off-targets in bold.

mm0_exon_Pop5_chr5_115238085 TGCGGTTCAAGCACAGGTAAGCAGCGCTGTCCCCGCGCCGTCCCCGCGACCCGGGCCGCGCTCACCCCGCTGTCGCCGCA
GGTACCTTCTGTGCGAG
mm4_exon_Agpat2_chr2_26603918 CTACAAGCGGCGCACATCGTGACACGCGCCCCTGGGACGGGAAGCCGCGTCGTCGGAGGCTCCGGACCGTGGGGACGGCG
GGG
CGGGCGGGGCAGGAAGGAAAGAACGCGAGG
mm4_intergenic_Banp|Gm22_chr8_122172637 CCATTCACCCTACAGCCTGGCCCCTGCTGTCCCTGCGACCCAGGGTCTAGTAAGTGCCCTGATGCCGCATCAAAGCCTGT
CATGTACACTCATGACCGGGACTGCACCTCTGAAGGTTCTGCAGCTGG
mm3_intron_Pmp22_chr11_63132535 GGCTGAAGGCTCTTCACCCCCCGGGGCGCAGGGAAGGCGCGGAACCCTGCGCGCGGGAAGCTCCGGAGCCGGGCAATCCT
CCAGCCCGCCTGCTCTCCCGCTCCCCCAGCCGCCAGCATCTAGGCCGAG
mm4_intron_Aff4_chr11_53351182 GGGAAATGGTTCGGGCCTAGCGGAGCGGGGACTGGAGGTGAGAAGCCGCNNNNNNNNNNNNNNNNNNNNNNNNNNNNNGT
CTGAGGCGCGGCCGCGCCGCGGGGAAGGCGAGGCTCCGGCCGCTGACGCTGGCGGAGACC
mm4_intergenic_Dpy19l1|Dpy19l2_chr9_24503221 CTCACTCTGGCCGGCGAGGGCGGGCCGGGAGGCGGGGACAGCGCGGGCCGTGTGGCCGCTGCCTACCGCGTGGCGGCCGA
GAGGCGGCGCCGGACTCCCGCGCCTCCGCCGTTGCCGCTCTCGGGTTGCCAGGG
mm4_intergenic_Kalrn|Ropn1_chr16_34573926 CTAGGCCGGCTTCCTGAGCCCCGCAGGAAAAGCCCGGGCCGAGGANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTC
TCCCGGCTTGCGGGGGACTCAGCGGCCGCCGCCGCAAGCTCCGCAAGCTCCGGGTC
mm3_intergenic_Chrdl2|Pold3_chr7_100042276 ACGGGCTTTGATTCGTGACTCCGGCGTGGACCCAGACGCTAATTAAAATCGCCTNNNNNNNNNNNNNNNNNNNNNNNNNA
CAAAGCCCGCTTTGACCTGGCGGCACAGGCCAGGCTTGTGGGACGTGTTTGTT
mm4_intron_Zfp398_chr6_47835952 GGTAAGGGCGCTGATGGACGGGCGTCTCTGTCACCCGGGCCTCGGGGGAGGCGAGGGAGCTGGGGACGGCAGCGGGCCTT
GTGGCCCGCGTGGGTCTGCGGCTGTCGTTTG
mm4_intergenic_Gzmk|BC067074_chr13_113293228 CGCGCAGGTAGAGCTCGGCGCGGGCCCCGGAGACGGCGCGGCGGGATGGGGCGGCCCGGGGCCTGCGGCCTCCTGTTGCT
CACCGCCTTTCTGGGG
mm4_exon_Rbm15_chr3_107332581 CCAGCTCGCTGATCTTCAGGGTCTTGTACTCCACGCCGTCCCCGCCGCCGGGCGCGGAGGAGGCGGCCCCGGAGGAACGT
GACTC
mm4_intron_6330403A02Rik_chr1_180483062 GACACATCCACGGGTCTCAGCCAGTGCCTACCTCTGCCCTGCCTGGTCTCCATCAGGCAGCCCAGGTCGCAGGGACCGCA
GGG
GTTAAAGGGGTCACAAGGGTCGCCCACACACACACTTCTTACCCAATCCCGGCAC
mm4_exon_Nudt9_chr5_104047125 TCAGTGACTGTCCGGTCCTCAGCCTGCCGCGCCGTCCCCGCACCCAGGTACCCCGATTCTAGGACGGGCTGCGGTTCCGA
GACCTCGGGTGGACCTGCGATCCCCTCCTTCCTTTT
mm4_intergenic_Gm13780|Phf21a_chr2_92183681 CCCAGCTCCAGTCCCTCTCGCCCGGCTCTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNGCTCCCGCGCGGGCGAGTGTGTCTGGGCCTG
mm3_intergenic_Elmod1|Tnfaip8l3_chr9_53975358 GTTTCCCTGGGAGCAAGACGCTGGCGGCTACTGAGCATGCTCAGTCGCGGAANNNNNNNNNNNNNNNNNNNNNNNNAGGA
GTGAGCTCAGGCCCCCGCGCCAGAATGTGGGAAGAAAGGGTTAAAACTGCGCACTTGGCC
mm4_intron_Ahi1_chr10_20952794 CTCTCTGTGAGTATCCCGCGGGCGCGCTGTGGCGGAGGCTCTGGCCTGCCCGGTCCCCGGGGGCGGCGAGGGGGCACCCG
GAGACTGTGGGATGCTC
mm4_exon_Atp8b1_chr18_64660909 GACTCCTCAGGCTCTCGTGGCTCGCCAAGGGCTCCCCGCCTTCCCCCCGCCCCGCGTCCGTGCGCCCTTCCTAGGCGGCA
GCCCTTTTCCCTCCGCTCTGCCCGACCGGACAAACTGGTTTCTCTGCTCCAGGG
mm3_exon_Gm26776/Ccdc3_chr2_5137874 TAGCCGTGCACCCAGCTCTCCGGAGCGCGTGCAGGCGAGCCGAGCGCCCCGTCCGCGGTTCTCGGGCAGGCGCTGCGGGC
TCCCCGGCTCCCCGCCGTCCCGGGCACCCGGGCGGGCCATGCGCCCGGGCTAGAGCGTAG
mm4_exon_B4galt3_chr1_171270381 CCAAGATGGCTGCAGCGATACCTGCCCGGTTGTCCGGGTGGCGGTGACGACGGGGAGCAGACCAGGTATGGTGGGCTGGG
GACCACGGAGAGAGAGAGT
mm4_intergenic_Ppm1h|Mirlet7i_chr10_122985563 CGGGGAGCTAACGGTTTCCGCGCCGGCCGTGCGGTCCCCGCGCCCCGGAAAACAAAGGGGATCCCCGGCCATTGTCCTCC
GCGGCGCTGAGCATCACCAGCACTAGCAAGGCAGTAGC
mm4_exon_Igf2bp3_chr6_49214416 CCGGGATCTTAGCGTCCTTGAAGACACTTTCCAGGTCCGCGGGGCCGGCGTGGTCGCTGAGGTTCCCGATGTACAATTTG
TTCATTGTGAAAAGGGGTCGTTTTAAAACCCGA
mm4_intron_Snx6_chr12_54795580 AAGAGAGACATTAGCGCCCANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNAGTTCGGCGCCGCCGTG
AACACCCACCATCATGGCTGCTTCGAGTCC
mm3_intron_Gm15608_chr10_82178505 CGGAATCCGCACCATGGTGAGCGGGACCGCCGTCCCCACGCGGGCGGAGCCGCGGGAGCCTGTTCGCCCTGGGCTGCGGG
GACGGCGAAG
AAACGTGGCACGCCAGGAGGTCTCTGCAGTCCCTGGAACTCACGCTC
mm4_intron_Usp2_chr9_44084846 AGGGTCATCGGCTTTGGGCGGGCCCTGCGACCTCGCCGGCGTCCGCGTGCCAACTCCNNNNNNNNNNNNNNNNNNNNNNN
NNNNNTGGTCCGGGTCCGGCTCTGCCCCTGCCCCTCCCCGGCGCTGACGTCGCAAAGCTG
mm4_intergenic_Gm12341|Abr_chr11_76562016 AAAGCTCTGGGCAGTTGAGGGGTGGTCAGCACGGCAGCAAGGCTGAGCCTCTGTGTTGGTTACATTCTGGAATCAAGGGT
CGCGGGGAGGGCTAGG
AGGCAGGGACAGAACAGGACCCCAGGA
mm4_intergenic_Ppp3r1|Wdr92/Ppp3r1_chr11_17159798 CTCCCCGAGATTCGCCGCCCCCGCCCTCCGCACGACCCTGGGCCTCCGGGCGCCCACAGAGCCGCCGCCCCCGGGCTCCT
AAGCCCGTGGGCCTCCGCGCAACCCCGGCCCCGTCGCCTGTCTGGCTCGTTCGGCCTTTT
mm4_intergenic_Gab1|Gm27048/Gm4891_chr8_80880608 GGGATCTGTAGTCAGCAGGCGCGCTGGGGTCCTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTATGGAAGGCGGCGGTCT
CCGGGAACGCAGCGCCACCCCAGTGTGAGAGCGGGAGTG
mm4_exon_Tbc1d4_chr14_101608150 TGCTCGAAGATGAACACCCCGGTGTTGGGCTGCACAGCCCCGGAGCCTGCGCCCCCGCCGACCCCGGCACCCGGCGCAGG
GACACAACGCAGAAAGGGGGCGCTGAGCACCAGGATCACCTC
mm4_intergenic_Gm16195|Srsf3_chr17_29032421 TAGGCGGAAGTGGAGGAGAGGGGGCGTCTGCGCGGCTCGCGGGGATGGCCGGGCCGAGCGCCGCATCTGCGGGACAAGCC
GGCACGCGGCGACGAGCTCGCGCCACCGCGCGTACCCCACCCACCTGACGGAAC
mm4_exon_Zfp664_chr5_124863026 ACAACCCATCCTTCAACCGGGCCTGGGCCGCGGGCATGGCGAGGAGGGAAGGGAAGGTCAGCGCGGTGCCCGCTGCGGCG
CAGGGCCCGGGTCGGGCGCGGGATGAGGCAGAAGAGGC
mm4_intergenic_Kcnj8|Abcc9_chr6_142571399 AGGTGCGATCCATAGTGCACGCGGGGAGGAGGGCTGGAAGATGAGGGTTGGGTTTGGCTTCCCCCCTCCTGCTGCCGGGC
CACGGGGAGGGCGGGG
TAGGAGCTGGGGATAGAGGTG
mm4_intergenic_Grin2b/Gm26653|Grin2b_chr6_136173328 CAGCAGGAGAGGAAGAGCAGGGGAGGAGGGGGAGAGTAAGAAAAATGGGTATTAATCACCGGCGACGCGGGGAGGGCGGG
G
ACATCAAAGGTCCTGGTCTTGATTTGGGTCTCACACCCAGGAACGTG
mm4_intergenic_Gm13012|Ece1_chr4_137897857 GCCCTCAGTTTTCCCATCCCAATCTCAAACATCTTTTCTCTTGTTATCGCTAATTCCAGAGAAGGTTGTCCCCCCGCCAT
CCCCCCGCCCCGTG
ACAATAACGGGTGCTTTTGCATTTGTGGATGGGGACA
mm4_exon_Arhgef16_chr4_154299870 GTCTTCCCCTTGGCTGACTCTCCAGTCCCCAGACTCACCTGTGCGCGCAACCGAGGGTCGGCCGGGAGGGTGTAGCCTGC
AGTGCCGCTGGGTCCCCGCGTCCCGGGGCCGGCCGGTCCCGCCCTCTCTACTTCCT
mm4_intron_Rala_chr13_17943812 CTTGAAAGCGAGGGAGGTCGGGGTGGAGTGCGGCGCAGGGACTNNNNNNNNNNNNNNNNNNNNCCCAGGTCGCGGGCACC
GCCCGG
GCACAGCCCTGGGAGCCACCGGCGCTATACAGGTGACCTT
mm3_intron_Sctr_chr1_120007340 CTGCTGCTCATCAATGCAGCGCACCCTGTGAGTACCCCGCCCCGTCCCCGCGCCCCCGGATGTGGACCCCACGGACCAGT
CCACGGTCCTCTATCCGGCCCTAGGAGTTACGGGGAGCAGCCGATCTCTCTTC
mm4_intergenic_Gm25266|Cradd_chr10_95136497 CCATGAAGTGGTGGTGTTTTCCTGTTTTAGTCACTAATATCCCCGGGTTGGGGGGGGGGCGGGGCGGGGAGGAGGATGCC
TGTGGGAGCAGAAA
mm4_intergenic_Usp15|Usp15/AC158802.1_chr10_123196796 GACCTGTCCGCACCTGTCACCAGCCGGCGACACCAGCCCACCGCTCCCGCCCAGCTCTCCGCCAGCCTNNNNNNNNNNNN
NNNNNNNNNACCCCGT
TCCCTACCAGGTGTCCCCTTTCCGGAGCGAGGTTTTGA
mm2_exon_Peli1_chr11_21091606 CTGGTGCCTGCGTTCAACCGGTGCTNNNNNNNNNNNNNNNNNNNNNNTGTCCGTGCGGCGGGCCGAGCTGGCGTTCCGGC
CACCAGCAGTCGTCCGGTGAG
mm4_exon_Drd4_chr7_141294210 TTTACTGGGCCACTTTCCGCGGCCTGCGGCGCTGGGAGGCAGCCCGGCACACCAAACTTCACANNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNN
NNNNNNNNTGTCGGACCCTACTCAGGGTCCCTTCTTCCCAGACTGC
mm4_exon_Fam211a_chr11_62648020 GACAGGTCAGCCTTCCTCACCCTAGCCTGAGGCCCCAAATTCAGCAGGAAACACAGAGGCACCTGCTGACACACCGCTCC
ATCCCCGCGAAACGGG
GACCCTGGGCGGTGCCTCCCGGCGCTCTCCTGTCACCC
mm4_exon_Prkcb_chr7_122289278 CTAGTCCCGCGCCTTTCCGGGCTTGCANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNACCTCTCGGGGCTCCCCCCAGTC
CCCGCGCGCGCAAGATGGCTGACCCGG
mm4_intergenic_Evx2|Hoxd13_chr2_74667870 GGGTGGCTGCAAAGTCAACTCCTGGGTCTGGGGGAGGGCGGGGATGCGAGGTGGGCCTCCCAAAGGGCTGAAGAGGGGTG
mm4_intergenic_Eif4h|Limk1_chr5_134639368 GTCTCCGCTCCTTGAGGAACCAGCGGGCCAGCGAGGAAGGACCCCGCAATATAACTCCTCCTCCCCCCGCCCGCCCCGCT
ACCGGGG
CTGTCCGCCGCGCTGCCTGATCGCAAGTC
mm4_exon_4930515G01Rik/Ankrd13a_chr5_114774982 TCGGTTCGCGAGCTGTCANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTCCGGACTCGGAGGCCCTTG

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_ozJbN0W08qlgEBn3RvZK.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.