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Contents:

PCR primers for off-targets of ACAGCGGGGTGAGCGCGGCC 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_115238103_F Primer3: not found at this Tm N.d. 1.00
mm0_exon_Pop5_chr5_115238103_R Primer3: not found at this Tm N.d. 1.00
mm4_intergenic_Gm24212|Tox3_chr8_90230423_F TCGTCGGCAGCGTCAACAAGATGTGAGCGTGCCT 60.2 0.73
mm4_intergenic_Gm24212|Tox3_chr8_90230423_R GTCTCGTGGGCTCGGCTGCCTTTTGTCCCACCTCA 60.1 0.73
mm3_exon_Ubald2_chr11_116434515_F TCGTCGGCAGCGTCGCAGTTCGAGGTGTGCAC 59.0 0.61
mm3_exon_Ubald2_chr11_116434515_R GTCTCGTGGGCTCGGCTGCGGACAAAATACGGGAC 59.2 0.61
mm4_intergenic_Gm22480|Gm23096_chr3_49624315_F TCGTCGGCAGCGTCTGTCTCCTTCTGCTGTTGGAC 59.9 0.52
mm4_intergenic_Gm22480|Gm23096_chr3_49624315_R GTCTCGTGGGCTCGGCCCTAGTCTGGAGCTGGACT 60.0 0.52
mm4_intron_Srsf11_chr3_158031820_F TCGTCGGCAGCGTCCCCATATCCCGGAAGTACGC 60.0 0.44
mm4_intron_Srsf11_chr3_158031820_R GTCTCGTGGGCTCGGTAGGTGGAGGGATTCTGCCT 59.6 0.44
mm4_exon_Tspan9_chr6_128143498_F Primer3: not found at this Tm N.d. 0.31
mm4_exon_Tspan9_chr6_128143498_R Primer3: not found at this Tm N.d. 0.31
mm4_intergenic_Rhob|Slc7a15_chr12_8519843_F Primer3: not found at this Tm N.d. 0.29
mm4_intergenic_Rhob|Slc7a15_chr12_8519843_R Primer3: not found at this Tm N.d. 0.29
mm4_exon_Tenm3_chr8_48555133_F TCGTCGGCAGCGTCAGAGAGAGCTCGGAGCCG 60.5 0.24
mm4_exon_Tenm3_chr8_48555133_R GTCTCGTGGGCTCGGCTCCAGACCCAGGCAAGTTG 60.6 0.24
mm3_intergenic_Gm15402|Tcf7l1_chr6_72724172_F Primer3: not found at this Tm N.d. 0.24
mm3_intergenic_Gm15402|Tcf7l1_chr6_72724172_R Primer3: not found at this Tm N.d. 0.24
mm3_intron_Akt1_chr12_112673951_F Primer3: not found at this Tm N.d. 0.22
mm3_intron_Akt1_chr12_112673951_R Primer3: not found at this Tm N.d. 0.22
mm4_intron_Tmem110_chr14_30854863_F Primer3: not found at this Tm N.d. 0.20
mm4_intron_Tmem110_chr14_30854863_R Primer3: not found at this Tm N.d. 0.20
mm4_exon_Tuba3a_chr6_125285952_F TCGTCGGCAGCGTCCTCAGAGGAGACGGAGGTCA 60.0 0.20
mm4_exon_Tuba3a_chr6_125285952_R GTCTCGTGGGCTCGGGCGTCTGAGCGGGTCTCC 62.5 0.20
mm4_intergenic_Gm7204|Dppa4_chr16_48236103_F Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Gm7204|Dppa4_chr16_48236103_R Primer3: not found at this Tm N.d. 0.19
mm4_exon_Dip2a_chr10_76345260_F Primer3: not found at this Tm N.d. 0.19
mm4_exon_Dip2a_chr10_76345260_R Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Gm833|Gm25779_chr4_152866704_F Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Gm833|Gm25779_chr4_152866704_R Primer3: not found at this Tm N.d. 0.18
mm4_exon_Shisa4_chr1_135374965_F TCGTCGGCAGCGTCCAAGCACCAGCAGGACGA 59.9 0.16
mm4_exon_Shisa4_chr1_135374965_R GTCTCGTGGGCTCGGGCTTCGTCCAGCTTTGTCCT 60.6 0.16
mm4_exon_Uchl3_chr14_101654097_F Primer3: not found at this Tm N.d. 0.14
mm4_exon_Uchl3_chr14_101654097_R Primer3: not found at this Tm N.d. 0.14
mm4_intron_Gnai3_chr3_108128899_F TCGTCGGCAGCGTCTGCTTATCTTCAATCTACCAAGGA 57.4 0.13
mm4_intron_Gnai3_chr3_108128899_R GTCTCGTGGGCTCGGCCAGGCTTGCCATCCTTACA 60.0 0.13
mm4_intergenic_Ppp1r13b|2010107E04Rik_chr12_111946421_F Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Ppp1r13b|2010107E04Rik_chr12_111946421_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Ppp1r13b|2010107E04Rik_chr12_111944098_F TCGTCGGCAGCGTCGCCCCACTCTGCAGAAAGAA 60.2 0.13
mm4_intergenic_Ppp1r13b|2010107E04Rik_chr12_111944098_R GTCTCGTGGGCTCGGTCATCTAGGTAGGGCTGGGG 59.8 0.13
mm4_intron_Mpdz_chr4_81442566_F Primer3: not found at this Tm N.d. 0.12
mm4_intron_Mpdz_chr4_81442566_R Primer3: not found at this Tm N.d. 0.12
mm4_exon_Gnb1_chr4_155491572_F Primer3: not found at this Tm N.d. 0.12
mm4_exon_Gnb1_chr4_155491572_R Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Gm22119|Gm6026_chrY_2577177_F Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Gm22119|Gm6026_chrY_2577177_R Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Gm16501|Rbmy/Gm10256_chrY_2733243_F TCGTCGGCAGCGTCCCTTGGGGATGTTTAGGCGT 60.0 0.12
mm4_intergenic_Gm16501|Rbmy/Gm10256_chrY_2733243_R GTCTCGTGGGCTCGGCTCTCCCAGTTGGAAACCCC 59.9 0.12
mm4_exon_Zc3h12a_chr4_125127751_F TCGTCGGCAGCGTCGGAGTCAGACCCAACGCAG 60.3 0.12
mm4_exon_Zc3h12a_chr4_125127751_R GTCTCGTGGGCTCGGCTACGAAGGTGCCGTCAGAG 60.1 0.12
mm3_exon_A730015C16Rik/Kti12_chr4_108848174_F Primer3: not found at this Tm N.d. 0.11
mm3_exon_A730015C16Rik/Kti12_chr4_108848174_R Primer3: not found at this Tm N.d. 0.11
mm4_exon_Tspan13_chr12_36042255_F TCGTCGGCAGCGTCGAATACTCAGGCGGGGAGGA 60.7 0.10
mm4_exon_Tspan13_chr12_36042255_R GTCTCGTGGGCTCGGGGCTTCTCGTGCTCCAAGAA 60.3 0.10
mm4_intron_Zdhhc17_chr10_111009730_F Primer3: not found at this Tm N.d. 0.09
mm4_intron_Zdhhc17_chr10_111009730_R Primer3: not found at this Tm N.d. 0.09
mm4_exon_Edar_chr10_58675590_F Primer3: not found at this Tm N.d. 0.08
mm4_exon_Edar_chr10_58675590_R Primer3: not found at this Tm N.d. 0.08
mm4_exon_Elavl4_chr4_110211483_F TCGTCGGCAGCGTCATCTTTGAGCCTGGTGGTGG 59.9 0.08
mm4_exon_Elavl4_chr4_110211483_R GTCTCGTGGGCTCGGAACAACCCCAGCCAGAAGTC 60.1 0.08
mm4_exon_Bmp2k_chr5_96997673_F Primer3: not found at this Tm N.d. 0.08
mm4_exon_Bmp2k_chr5_96997673_R Primer3: not found at this Tm N.d. 0.08
mm4_exon_C1ql3_chr2_13010244_F TCGTCGGCAGCGTCTCTCCTCCCTCTTGTCAGCA 59.8 0.07
mm4_exon_C1ql3_chr2_13010244_R GTCTCGTGGGCTCGGCATGTGGGCTGATCTCTGCA 60.1 0.07
mm3_exon_Galc_chr12_98259119_F TCGTCGGCAGCGTCCACCGCACTCCTGCCTCC 62.7 0.07
mm3_exon_Galc_chr12_98259119_R GTCTCGTGGGCTCGGATTGTTGTGTGCGCTGCTAG 59.4 0.07
mm4_intergenic_Cd47|Gm8824_chr16_50017123_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Cd47|Gm8824_chr16_50017123_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Chrdl2|Pold3_chr7_100042164_F TCGTCGGCAGCGTCCAGCGCAGGGATCACTCTAG 59.9 0.06
mm4_intergenic_Chrdl2|Pold3_chr7_100042164_R GTCTCGTGGGCTCGGAGTCACGAATCAAAGCCCGT 59.9 0.06
mm3_intergenic_Gm17111/Dync1h1|Dync1h1_chr12_110665429_F TCGTCGGCAGCGTCAGGCAGCTCCTGTGACTTTC 59.9 0.06
mm3_intergenic_Gm17111/Dync1h1|Dync1h1_chr12_110665429_R GTCTCGTGGGCTCGGGGTGCCTGCTTAGAGGTGTT 59.9 0.06
mm4_intergenic_Nrd1/Osbpl9|Osbpl9_chr4_109062376_F TCGTCGGCAGCGTCGCCATCCTCGTGAAAGAGCT 60.1 0.06
mm4_intergenic_Nrd1/Osbpl9|Osbpl9_chr4_109062376_R GTCTCGTGGGCTCGGGCTCGTCCATGCCAATGAAG 59.6 0.06
mm4_intron_Ldha_chr7_46850359_F TCGTCGGCAGCGTCCAAGCTGCTGATCGTCTCCA 60.1 0.06
mm4_intron_Ldha_chr7_46850359_R GTCTCGTGGGCTCGGAGTGTAGCTAACGCACCGTG 60.3 0.06
mm4_intergenic_B230319C09Rik|Dguok_chr6_83475204_F TCGTCGGCAGCGTCCCTGCTGCTACATTTCCCCA 60.0 0.05
mm4_intergenic_B230319C09Rik|Dguok_chr6_83475204_R GTCTCGTGGGCTCGGCCCAACTGAGCCTAAGGAGC 60.1 0.05
mm4_exon_Epha5_chr5_84417175_F TCGTCGGCAGCGTCGCTGGCAGAGGGAAGTCAAA 60.2 0.05
mm4_exon_Epha5_chr5_84417175_R GTCTCGTGGGCTCGGCTCTGCTGCGCTCTCGGG 62.8 0.05
mm4_exon_Gpr88_chr3_116251657_F TCGTCGGCAGCGTCGAACTCCTCGTTCCTCCACG 60.1 0.05
mm4_exon_Gpr88_chr3_116251657_R GTCTCGTGGGCTCGGCTGGTGTGGGTGAGCCTG 59.9 0.05
mm4_intron_Tex261_chr6_83775616_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Tex261_chr6_83775616_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Rnf214|Rnf214/Pcsk7_chr9_45906186_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Rnf214|Rnf214/Pcsk7_chr9_45906186_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Kcnq3|RP24-108J19.1_chr15_66286707_F TCGTCGGCAGCGTCTTTACAAGCCCTGTGCCAG 57.9 0.04
mm4_intergenic_Kcnq3|RP24-108J19.1_chr15_66286707_R GTCTCGTGGGCTCGGCTGCTGCTGCTACTGCCG 61.1 0.04
mm4_exon_Plk1_chr7_122169773_F TCGTCGGCAGCGTCTTGAAGGGGTTGCTGTGTGT 60.0 0.04
mm4_exon_Plk1_chr7_122169773_R GTCTCGTGGGCTCGGGCTAAGGAAAGAGCAGTCCCA 59.7 0.04
mm4_intron_Cxcr4_chr1_128591027_F TCGTCGGCAGCGTCTACTGGAGTCCGAGGTAGGC 60.1 0.04
mm4_intron_Cxcr4_chr1_128591027_R GTCTCGTGGGCTCGGCCCTAGAACCGCTGTGTGTT 59.9 0.04
mm4_intron_Egln1_chr8_124922049_F TCGTCGGCAGCGTCAATGTCTGCCCCATGTGGAG 60.0 0.04
mm4_intron_Egln1_chr8_124922049_R GTCTCGTGGGCTCGGACACTCAGGCTATGCTCAGC 59.8 0.04
mm4_intergenic_Serpine3|Gm25162_chr14_62666577_F TCGTCGGCAGCGTCACGGCTGGTCAGAAACTTGT 59.8 0.03
mm4_intergenic_Serpine3|Gm25162_chr14_62666577_R GTCTCGTGGGCTCGGCTCCAACCTGCTCTGTGGAA 59.6 0.03
mm4_intergenic_Sgsm1|Aym1_chr5_113346606_F TCGTCGGCAGCGTCCTTCGGGACAGTGAGGTGTC 60.0 0.03
mm4_intergenic_Sgsm1|Aym1_chr5_113346606_R GTCTCGTGGGCTCGGCTCCCAAGGTCTGGTCACAC 59.9 0.03
mm4_intergenic_Gm13821|mmu-mir-6240_chr5_114847585_F TCGTCGGCAGCGTCAGCTCCTGGTCACACCTAGA 59.5 0.03
mm4_intergenic_Gm13821|mmu-mir-6240_chr5_114847585_R GTCTCGTGGGCTCGGACGTGTGCTGCTTTGGGATA 59.9 0.03
mm4_intergenic_Camsap2|9230116N13Rik_chr1_136346148_F TCGTCGGCAGCGTCTGGGAACACTGAGCATGCG 60.6 0.03
mm4_intergenic_Camsap2|9230116N13Rik_chr1_136346148_R GTCTCGTGGGCTCGGATTAATGGATAGGGGCGCGG 60.0 0.03
mm4_exon_Chst11_chr10_82985833_F TCGTCGGCAGCGTCCTCTGTTCTGAGCCTCCCC 59.0 0.03
mm4_exon_Chst11_chr10_82985833_R GTCTCGTGGGCTCGGTCCTCATCACTTCCAGCAGC 59.7 0.03
mm4_intergenic_Sbno1|Sbno1/AC127339.1_chr5_124425283_F TCGTCGGCAGCGTCCGGACAGCAGGAGGAAACAG 60.6 0.03
mm4_intergenic_Sbno1|Sbno1/AC127339.1_chr5_124425283_R GTCTCGTGGGCTCGGCACACCACGGCTCCTCAG 60.0 0.03
mm4_intergenic_Gm26214|Zcchc2_chr1_105990156_F TCGTCGGCAGCGTCGGCACGGTCAGATCTCTCAA 59.4 0.02
mm4_intergenic_Gm26214|Zcchc2_chr1_105990156_R GTCTCGTGGGCTCGGTGGGTTTGGGACTTTCGGAC 60.1 0.02
mm4_intergenic_Gm14513|Gm26314_chrX_11723325_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm14513|Gm26314_chrX_11723325_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Atp9a_chr2_168742022_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Atp9a_chr2_168742022_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Pitx1_chr13_55831403_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Pitx1_chr13_55831403_R Primer3: not found at this Tm N.d. 0.02
mm3_intron_1810041L15Rik_chr15_84446500_F Primer3: not found at this Tm N.d. 0.01
mm3_intron_1810041L15Rik_chr15_84446500_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Peli2_chr14_48121053_F TCGTCGGCAGCGTCGATAGCTCCGGGGAAGCC 59.2 0.01
mm4_exon_Peli2_chr14_48121053_R GTCTCGTGGGCTCGGCGCCACTACACTCCCATCC 59.8 0.01
mm4_intron_Coro2b_chr9_62477300_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Coro2b_chr9_62477300_R Primer3: not found at this Tm N.d. 0.01
mm4_intron_Calm1_chr12_100199947_F TCGTCGGCAGCGTCACCTCCTCTGAGTTTTGGGC 59.5 0.01
mm4_intron_Calm1_chr12_100199947_R GTCTCGTGGGCTCGGGCGTTGGATGCAGTGGGT 60.6 0.01
mm4_intron_Ptk2_chr15_73422802_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Ptk2_chr15_73422802_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Tll1_chr8_64205753_F Primer3: not found at this Tm N.d. 0.01
mm4_exon_Tll1_chr8_64205753_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Akap8l_chr17_32332463_F TCGTCGGCAGCGTCGGCTCAAATTCCAGGCAAGC 60.1 0.00
mm4_exon_Akap8l_chr17_32332463_R GTCTCGTGGGCTCGGTGCAGCCTGTGACCTTTTCA 60.1 0.00
mm4_intron_Nfic_chr10_81430558_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Nfic_chr10_81430558_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tprg_chr16_25388684_F TCGTCGGCAGCGTCAGTGTGTTGTGATCTCTGGTCA 59.3 0.00
mm4_intron_Tprg_chr16_25388684_R GTCTCGTGGGCTCGGACACAGAATAGTAAATGCCTGCA 58.4 0.00
mm4_intron_Prkx_chrX_77791029_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Prkx_chrX_77791029_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Lrp5_chr19_3628694_F TCGTCGGCAGCGTCCCCTTGTCCCTGACCTGTTT 59.5 0.00
mm4_intron_Lrp5_chr19_3628694_R GTCTCGTGGGCTCGGAAACTTCTGCCCTCTGCCTC 59.9 0.00
mm4_intron_Cenpm_chr15_82244522_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Cenpm_chr15_82244522_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_C4a_chr17_34810414_F TCGTCGGCAGCGTCGCAAGACATGAGGGCCTCTT 60.0 0.00
mm4_intron_C4a_chr17_34810414_R GTCTCGTGGGCTCGGCAGCGCTGTCCTGTATGACT 59.8 0.00
mm4_intergenic_Pnpla3|Samm50_chr15_84190812_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Pnpla3|Samm50_chr15_84190812_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm26089|Fat1_chr8_44946281_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm26089|Fat1_chr8_44946281_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm11468|Gm14268_chr2_166307903_F TCGTCGGCAGCGTCGCTCAAGGCCATTTTGTGATT 57.6 0.00
mm4_intergenic_Gm11468|Gm14268_chr2_166307903_R GTCTCGTGGGCTCGGATGAGAGCTCCCCAGAGTGT 59.9 0.00
mm4_intergenic_1700041M05Rik|Grik3_chr4_125425605_F TCGTCGGCAGCGTCATGCCTCTCATTTGCAGCCT 60.0 0.00
mm4_intergenic_1700041M05Rik|Grik3_chr4_125425605_R GTCTCGTGGGCTCGGTGCATAGTCTCAGGACAGTGC 59.7 0.00
mm4_exon_Lipe_chr7_25388269_F TCGTCGGCAGCGTCCTCCTGGTCGGTTGATGGTC 60.1 0.00
mm4_exon_Lipe_chr7_25388269_R GTCTCGTGGGCTCGGCAACCTAGCAGAGCTGGAGG 59.8 0.00
mm3_intron_Dennd4c_chr4_86762418_F Primer3: not found at this Tm N.d. 0.00
mm3_intron_Dennd4c_chr4_86762418_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.

mm4_intergenic_Gm24212|Tox3_chr8_90230423 AACAAGATGTGAGCGTGCCTAATAAAAAGGGCCCACCCTTGCTGATTTATAGCCATCTAATTACCTGACAAATATCCAGG
CTGCGTTCACCCTGCTGG
GTAGCAGTATGTCAATATGAGGTGGGACAAAAGGCAG
mm3_exon_Ubald2_chr11_116434515 GCAGTTCGAGGTGTGCACCNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNCCAGGCCTCGCTCACGCCGCTGCGTCCCGTATTTTGTCCGCAG
mm4_intergenic_Gm22480|Gm23096_chr3_49624315 TGTCTCCTTCTGCTGTTGGACTCAGAGTCTGTCTGGCTGTAACCTGTATAGAGGCAGAGGATACAGGGAGGTGAACACGG
CCTGG
ATTCCCTGCTCAATTCACTGGAGCCTAAGTCCAGCTCCAGACTAGGG
mm4_intron_Srsf11_chr3_158031820 CCCATATCCCGGAAGTACGCGTCCAGTTCGCGCTCCCGCAACTGTCCCGGCTGCGCCCACCCAGCTGAGCCACCTCAGAA
ATGGTTCCTGCTTACTCCACATAGCACCAGAAGCATTTTCAGGCAGAATCCCTCCACCTA
mm4_exon_Tenm3_chr8_48555133 AGAGAGAGCTCGGAGCCGCGCGTGGAACCGCGCGAGGAACCGCCCAGGCCGCGCTCACCCACCTTGCCGCCGCGCGCCCT
CGCCGTGGCCGTGCGGAGCGGAGCCCGGCGCTGCCAACTTGCCTGGGTCTGGAG
mm4_exon_Tuba3a_chr6_125285952 CTCAGAGGAGACGGAGGTCAACCTCCCGTGGGGCCGACCCCCCCAGAGGCCCCGAGGCCAACCACCCCGGCCTGGGACTG
CTCCCACCACCCCGGCCGCTCTCACCATGCTGAACTCCGGAGACCCGCTCAGACGC
mm4_exon_Shisa4_chr1_135374965 CAAGCACCAGCAGGACGACGGCTGCGANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNATGGCGAGAGGGGGCTGGGCG
CGGCCAGG
ACGGCCTGGAAAAGGGGCCGCAGGACAAAGCTGGACGAAGC
mm4_intron_Gnai3_chr3_108128899 TGCTTATCTTCAATCTACCAAGGATAATAGGCTATAAGAACTGGAACAGCTAGAGCCTACAGCGGAGGGAACGCGGCTCG
G
AGTAGGAGGAACTGAAGATGAGCCTGCAGACTGTAAGGATGGCAAGCCTGG
mm4_intergenic_Ppp1r13b|2010107E04Rik_chr12_111944098 GCCCCACTCTGCAGAAAGAAGGTCTCCCTAGGAGACAGCAACAGATGCCACAGACTGGCCCTGGCCATGCTCATCCCGCT
TT
CAGGGACCCACGAGTCCCTCCTCCCCCAGCCCTACCTAGATGA
mm4_intergenic_Gm16501|Rbmy/Gm10256_chrY_2733243 CCTTGGGGATGTTTAGGCGTGTCCTGCAAGGGAATGGGCGTGATCTATGCTCTGCAGGCATGTCCTAAGTTGTGGGTGTG
TTCTCAGCGTGGTGGGCGTGGCCTGGACGCTTTGGGGTTTCCAACTGGGAGAG
mm4_exon_Zc3h12a_chr4_125127751 GGAGTCAGACCCAACGCAGCGCAGCACCCGTTCCACCCGGTAGCACTCACCCCGCTGAGCGCCGGACGTCGTCTGTAAGG
TGTCCATGGAGCCGAGTCTCGGGCTCTGACGGCACCTTCGTAG
mm4_exon_Tspan13_chr12_36042255 GAATACTCAGGCGGGGAGGAGGCCGACGGCCCCCGCAGGTGCCACCGGGCGCGGGCTACTCACGGTATACAGCAGGTTGA
GCGCGCACAGG
CAGTTCTTGGAGCACGAGAAGCC
mm4_exon_Elavl4_chr4_110211483 ATCTTTGAGCCTGGTGGTGGAGAGGGCCAGGGTATCGCCTGTTGGGGGACTGGTACAGCTGGGAGAGCAGGGCCTGGCTG
GACTTCTGGCTGGGGTTGTT
mm4_exon_C1ql3_chr2_13010244 TCTCCTCCCTCTTGTCAGCACCTTCTCCGCCCTCCCCACTGCCTGCCCGCGCTCACCTGGTTGTTTTTGCAGAGATCAGC
CCACATG
mm3_exon_Galc_chr12_98259119 CACCGCACTCCTGCCTCCCGCCAGCCCGCGCTCACCCCGCCGCCGCTGACTGCGCCGATGCCATCGAACTCCCGGCCCAG
CCCGTCGGAGTCGTCTAGCACGTAGGCGCCACCGGGCACTAGCAGCGCACACAACAAT
mm4_intergenic_Chrdl2|Pold3_chr7_100042164 CAGCGCAGGGATCACTCTAGAGGCAAGACTCGAACTCCTCCACTGCCCGCTGCTGCCAGCAGGGTGCTCGCGGCCCGGGA
CAGGGGGCGGGGGTCGGGGCACCGGCGCAGAAGAACACGGGCTTTGATTCGTGACT
mm3_intergenic_Gm17111/Dync1h1|Dync1h1_chr12_110665429 AGGCAGCTCCTGTGACTTTCAGGGTTAATGCAGTGAAGAGGACAGAGGGGTGGGCGGGGCCTGGTCCAGACACACTGCAC
GCTGCAACACCTCTAAGCAGGCACC
mm4_intergenic_Nrd1/Osbpl9|Osbpl9_chr4_109062376 GCCATCCTCGTGAAAGAGCTACAAAAGGAAGAGAGGGTTTGCTTAGGCAGTGCAGCTTTATTCCATGCCACGCTCATCCC
ACTGT
TAGGCAGGAACCCAGGTCCCCCTCTGCATCCTTCATTGGCATGGACGAGC
mm4_intron_Ldha_chr7_46850359 CAAGCTGCTGATCGTCTCCAATCCAGGTGAGCAGCCAGCCTGCGCTCGCCCTGCTGTGGCACGGTGCGTTAGCTACACT
mm4_intergenic_B230319C09Rik|Dguok_chr6_83475204 CCTGCTGCTACATTTCCCCAGTCGGGGGCCCAGGCCACTCCCACCCCGCAGTCTCCTCCTCTTGAGGTACTTCATATTCT
AAAGCTGCTCCTTAGGCTCAGTTGGG
mm4_exon_Epha5_chr5_84417175 GCTGGCAGAGGGAAGTCAAATCGCTGCGGGCTGAGCTCGGCCGGGGGCGGTGCGACCGGCGGAGCGGGCTGGAGCGCGGG
CCCGAGAGCGCAGCAGAG
mm4_exon_Gpr88_chr3_116251657 GAACTCCTCGTTCCTCCACGTGTAGAGCAGCGGGTTGAGCGCCGACAGGGCGCAGCACAGGAGCCAGCTGGCCGCCTGCA
CGCCCCAGGGCACCGGGAGGGAGAAGCCACTGGCCAGGCTCACCCACACCAG
mm4_intergenic_Kcnq3|RP24-108J19.1_chr15_66286707 TTTACAAGCCCTGTGCCAGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNGCTGCAGCGGCAGTAGCAGCAGCAG
mm4_exon_Plk1_chr7_122169773 TTGAAGGGGTTGCTGTGTGTAAGTTATTTTGTACATGTCTGGGTGTGGGTTTACATCTGCTTCCCTGCCCTCGCTCAGCC
CACTGT
ATGAATTGTATAAATGTTTCTATTAAATTGGGACTGCTCTTTCCTTAGC
mm4_intron_Cxcr4_chr1_128591027 TACTGGAGTCCGAGGTAGGCAGCAGGAGTGGTCCAGACCGTGCGCACCCCGATGTCCGAGTCACCCACCTAGGGGCTGGC
ACCGTTGCCCATCATACCCAGGTCTACGACCAACACACAGCGGTTCTAGGG
mm4_intron_Egln1_chr8_124922049 AATGTCTGCCCCATGTGGAGGGGGCCTGGATCACCTGCCCGGCCAGCCTGGCAGCGATCGCCCTGCTGTGGCCCAGCTGA
GCATAGCCTGAGTGT
mm4_intergenic_Serpine3|Gm25162_chr14_62666577 ACGGCTGGTCAGAAACTTGTCTTATGCACATGAGGAGAACTCTGAAGGCAGCAACTCTGTCTGGGAGAGCAATACATGTA
GACATGGGGGTGAGTGCCGCCTGGTTCTGTGATTCCACAGAGCAGGTTGGAG
mm4_intergenic_Sgsm1|Aym1_chr5_113346606 CTTCGGGACAGTGAGGTGTCAGCATCCAGGCCCGGAGGGATTCCCAGCAGGCCCAGTTTAACAGAGGGGTGACCAGGGCC
CGG
AAAACTCTTGTCTAGGTGTGACCAGACCTTGGGAG
mm4_intergenic_Gm13821|mmu-mir-6240_chr5_114847585 AGCTCCTGGTCACACCTAGACAAGAGTTTTCCGGGCCCTGGTCACCCCTCTGTTAAACTGTGGGCCTGCTGGGAATCCCT
CCAGGCCTGGATGCTGACACCTCACTATCCCAAAGCAGCACACGT
mm4_intergenic_Camsap2|9230116N13Rik_chr1_136346148 TGGGAACACTGAGCATGCGCCTNNNNNNNNNNNNNNNNNNNNNNNNNNTGTCTTGCGCCGCCATGCGCACTGAGCATGCC
CAGCCTCGCTGCCGCCCGCGATGGGCTCCGCGCCCCTATCCATTAAT
mm4_exon_Chst11_chr10_82985833 CTCTGTTCTGAGCCTCCCCGGTGCTGGTACCCCCGCGGCGGGGTCCCCGCTTCTGCGCTCGGGACGCGCCTCCCGGACAG
CCGGGTCCCCGCGGCCAGG
ACAAAGCCATGAAGCCGGCGCTGCTGGAAGTGATGAGGA
mm4_intergenic_Sbno1|Sbno1/AC127339.1_chr5_124425283 CGGACAGCAGGAGGAAACAGAGCAGCGGGGCGAGCGAGGCGCGGACGGCCATAGAGTGGCCGCCTCCCCGCCACCCCCCA
CAGACGGCCCCACACGGACCCTGAGGAGCCGTGGTGTG
mm4_intergenic_Gm26214|Zcchc2_chr1_105990156 GGCACGGTCAGATCTCTCAAGAAAAACTAAGTCGATGTCCCTCCTCCCACGCGAACCTCGCCCACTTCTGCCGCTCCGGG
CCAAGCTCACCTCGCCGT
GGTCCGAAAGTCCCAAACCCA
mm4_exon_Peli2_chr14_48121053 GATAGCTCCGGGGAAGCCGGCGGGCGACGCGCACAGCCACGACGGAGCAGCCGCGGGACTGGCCGCCCAGCGCCCCGTTC
GCCGCCGTGCCGTTCCCCGGCGCGCTCACCCCGTTTTCGGGATGGGAGTGTAGTGGCG
mm4_intron_Calm1_chr12_100199947 ACCTCCTCTGAGTTTTGGGCGCCTCTGGGGCTGGACTCTACAGTGGGCTGTGCGCGGGCTGGAGCCGCTGCCACAGCTGC
ACCGGGGCCTTCCGCATCCACCCCCTCCGCCAGCCCCGCACCCACTGCATCCAACGC
mm4_exon_Akap8l_chr17_32332463 GGCTCAAATTCCAGGCAAGCAGCCCCGGGCCCAGCTCACCCTGCGGTTTCGGTTGTGATCCATCGTCTTGAGATGCTTTT
GGATGATTCCAAACTGCATGGGGATGAAAAGGTCACAGGCTGCA
mm4_intron_Tprg_chr16_25388684 AGTGTGTTGTGATCTCTGGTCAAAGACAGAATACTTCTTTTGCTAAAGAATAACTATGTCTCTAGGATGGAGAGAGGGGT
GAGGGCTGCCTGG
TTGCAGGCATTTACTATTCTGTGT
mm4_intron_Lrp5_chr19_3628694 CCCTTGTCCCTGACCTGTTTGCTAGAGTCCAGAACAGCGGGAGGAGGGCGGCAGGGAGCCTTCTAGAAACAGAAGCAAAG
CTCCCATATGCTCAGGATGGTGACCCCAGAAAAATTTGGAGGCAGAGGGCAGAAGTTT
mm4_intron_C4a_chr17_34810414 GCAAGACATGAGGGCCTCTTGGAGCCCAGGCCTCCCCCACCCCGCGGTGTCCCCGATGAGAGGGCTAACCCCCACAATCA
TGCTTGCCAAAGCATCCCAAGTGCTCACCAGGGCTGTAGTAGTCATACAGGACAGCGCTG
mm4_intergenic_Gm11468|Gm14268_chr2_166307903 GCTCAAGGCCATTTTGTGATTAAATGAAAGGGGGAGGGGGTTGGGAGCTTCCCACAGCGGAGTGAGGACGGACAGGAGAG
GAGAGACACACTCTGGGGAGCTCTCAT
mm4_intergenic_1700041M05Rik|Grik3_chr4_125425605 ATGCCTCTCATTTGCAGCCTCTGCAGTGGTATAAGAGAGGCTATGGGAGTTAAGGTGAGAATCAGATAGAGTTTGTGTCT
CCTGGCAGACCTCACCCAGCTGTGGACGCTGGCACTGTCCTGAGACTATGCA
mm4_exon_Lipe_chr7_25388269 CTCCTGGTCGGTTGATGGTCAGCAGGCGCTGGGCATAGTAGGCCATAGCACGGAGCTGGGTGAGGGCGGCCAGGTAGGCC
TCCAGCTCTGCTAGGTTG

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.