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Contents:

PCR primers for off-targets of CCCCGGACTCTCCAGAGGGG 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.

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
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52313821_F TCGTCGGCAGCGTCAGGGAGCCTGGGATTAGGAG 60.1 1.00
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52313821_R GTCTCGTGGGCTCGGCATGTCCTTTCGGCTCTCCA 59.7 1.00
mm4_intron_Csmd2_chr4_128299768_F Primer3: not found at this Tm N.d. 0.75
mm4_intron_Csmd2_chr4_128299768_R Primer3: not found at this Tm N.d. 0.75
mm4_intron_Pitpnb_chr5_111380806_F TCGTCGGCAGCGTCAGTTGTGCCAGCACTTCAGT 60.1 0.61
mm4_intron_Pitpnb_chr5_111380806_R GTCTCGTGGGCTCGGTGCTTGAGAGAAGTGGTCAGG 59.6 0.61
mm2_intergenic_Kctd3|Kcnk2_chr1_189088957_F Primer3: not found at this Tm N.d. 0.59
mm2_intergenic_Kctd3|Kcnk2_chr1_189088957_R Primer3: not found at this Tm N.d. 0.59
mm3_exon_Krba1_chr6_48416008_F Primer3: not found at this Tm N.d. 0.55
mm3_exon_Krba1_chr6_48416008_R Primer3: not found at this Tm N.d. 0.55
mm4_intron_Uty_chrY_1201504_F Primer3: not found at this Tm N.d. 0.53
mm4_intron_Uty_chrY_1201504_R Primer3: not found at this Tm N.d. 0.53
mm4_intron_Mgam_chr6_40634046_F Primer3: not found at this Tm N.d. 0.53
mm4_intron_Mgam_chr6_40634046_R Primer3: not found at this Tm N.d. 0.53
mm4_intergenic_Gm26099|Gm24809_chrX_37067034_F Primer3: not found at this Tm N.d. 0.53
mm4_intergenic_Gm26099|Gm24809_chrX_37067034_R Primer3: not found at this Tm N.d. 0.53
mm4_intergenic_Gm26076|Gm25519_chr3_111465444_F Primer3: not found at this Tm N.d. 0.53
mm4_intergenic_Gm26076|Gm25519_chr3_111465444_R Primer3: not found at this Tm N.d. 0.53
mm4_intergenic_Gm26341|Amd2_chr10_35532437_F Primer3: not found at this Tm N.d. 0.50
mm4_intergenic_Gm26341|Amd2_chr10_35532437_R Primer3: not found at this Tm N.d. 0.50
mm3_intergenic_1700013D24Rik|Pex5_chr6_124392546_F Primer3: not found at this Tm N.d. 0.48
mm3_intergenic_1700013D24Rik|Pex5_chr6_124392546_R Primer3: not found at this Tm N.d. 0.48
mm4_intron_Ctnnd2_chr15_30302354_F TCGTCGGCAGCGTCGAACGGACCTGCTTCACTGA 59.9 0.45
mm4_intron_Ctnnd2_chr15_30302354_R GTCTCGTGGGCTCGGAGCGGTTCTGAAACAAGAAAGC 59.9 0.45
mm2_intron_Sufu_chr19_46477778_F Primer3: not found at this Tm N.d. 0.44
mm2_intron_Sufu_chr19_46477778_R Primer3: not found at this Tm N.d. 0.44
mm4_intergenic_BC017158|Rgs10_chr7_128307285_F TCGTCGGCAGCGTCTGGACATCTCTCCCACCTGG 60.6 0.43
mm4_intergenic_BC017158|Rgs10_chr7_128307285_R GTCTCGTGGGCTCGGACCTTGGGGTGTTCCAGAGA 60.4 0.43
mm4_intron_Sema4f_chr6_82931569_F Primer3: not found at this Tm N.d. 0.39
mm4_intron_Sema4f_chr6_82931569_R Primer3: not found at this Tm N.d. 0.39
mm4_intron_Ppa1_chr10_61653066_F TCGTCGGCAGCGTCCCATAAGACTCCGCCGTCTC 59.9 0.39
mm4_intron_Ppa1_chr10_61653066_R GTCTCGTGGGCTCGGGTCAGCCAAGGGTTACCACA 59.8 0.39
mm4_intergenic_6430571L13Rik|Cacna2d2_chr9_107366351_F Primer3: not found at this Tm N.d. 0.39
mm4_intergenic_6430571L13Rik|Cacna2d2_chr9_107366351_R Primer3: not found at this Tm N.d. 0.39
mm4_intergenic_Gm12066|Gm12068_chr11_24349134_F TCGTCGGCAGCGTCGCGTATCAGAGGCTGAGTGT 59.5 0.36
mm4_intergenic_Gm12066|Gm12068_chr11_24349134_R GTCTCGTGGGCTCGGTGTCATTAGCAGGCACCCTC 59.7 0.36
mm3_intergenic_Rora|F830001A07Rik_chr9_68823588_F TCGTCGGCAGCGTCTATCGTGGTGGAGCTGATGC 59.8 0.35
mm3_intergenic_Rora|F830001A07Rik_chr9_68823588_R GTCTCGTGGGCTCGGATGACTGACCATGCCTGACC 59.7 0.35
mm4_intron_Atg2b_chr12_105642191_F TCGTCGGCAGCGTCACCATTAGCTACCAACAAACCCA 60.1 0.34
mm4_intron_Atg2b_chr12_105642191_R GTCTCGTGGGCTCGGCCAGCAGGGTTTTAAGGAACC 59.1 0.34
mm4_exon_Tor1aip2/Tor1aip1_chr1_156035585_F Primer3: not found at this Tm N.d. 0.33
mm4_exon_Tor1aip2/Tor1aip1_chr1_156035585_R Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Gm22674|Cdyl_chr13_35541958_F Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Gm22674|Cdyl_chr13_35541958_R Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Raxos1/Rax|Cplx4_chr18_65941747_F Primer3: not found at this Tm N.d. 0.32
mm4_intergenic_Raxos1/Rax|Cplx4_chr18_65941747_R Primer3: not found at this Tm N.d. 0.32
mm4_exon_Syt13_chr2_92954521_F Primer3: not found at this Tm N.d. 0.28
mm4_exon_Syt13_chr2_92954521_R Primer3: not found at this Tm N.d. 0.28
mm4_intergenic_Syce1l|Adamts18_chr8_113688097_F TCGTCGGCAGCGTCGATAGACAGGACAGAGCGGC 59.9 0.28
mm4_intergenic_Syce1l|Adamts18_chr8_113688097_R GTCTCGTGGGCTCGGAGAGGGAGATACAGCCTGCA 60.0 0.28
mm3_intron_Plekha7_chr7_116288629_F TCGTCGGCAGCGTCGGGATGACTACCTGGGGACT 60.0 0.28
mm3_intron_Plekha7_chr7_116288629_R GTCTCGTGGGCTCGGGCCAGCAACTTCCTCTCACT 59.9 0.28
mm4_intergenic_Peli2|Gm23881_chr14_48361056_F TCGTCGGCAGCGTCTCTTTCCACGGTGTGCTGTT 60.1 0.28
mm4_intergenic_Peli2|Gm23881_chr14_48361056_R GTCTCGTGGGCTCGGCATTTCTCGAACCCCAGGCA 60.3 0.28
mm4_intergenic_Gm16494|Gm4945_chr17_47033724_F Primer3: not found at this Tm N.d. 0.27
mm4_intergenic_Gm16494|Gm4945_chr17_47033724_R Primer3: not found at this Tm N.d. 0.27
mm4_exon_Elavl4_chr4_110211633_F TCGTCGGCAGCGTCGGCTTCTTCTGCCTCAATCCT 60.0 0.26
mm4_exon_Elavl4_chr4_110211633_R GTCTCGTGGGCTCGGAGGCAAAATATCCACAGGGCA 59.9 0.26
mm0_exon_Nynrin/Gm26751_chr14_55871430_F Primer3: not found at this Tm N.d. 0.25
mm0_exon_Nynrin/Gm26751_chr14_55871430_R Primer3: not found at this Tm N.d. 0.25
mm4_intron_Ryr1_chr7_29078196_F Primer3: not found at this Tm N.d. 0.25
mm4_intron_Ryr1_chr7_29078196_R Primer3: not found at this Tm N.d. 0.25
mm4_intergenic_Gm8730|Cdh5_chr8_103672674_F Primer3: not found at this Tm N.d. 0.24
mm4_intergenic_Gm8730|Cdh5_chr8_103672674_R Primer3: not found at this Tm N.d. 0.24
mm4_intron_Zfp64_chr2_168911038_F TCGTCGGCAGCGTCGGGGCAAGTAGCTGAAGATGA 59.7 0.23
mm4_intron_Zfp64_chr2_168911038_R GTCTCGTGGGCTCGGGTACCTTAGGTGGAGTGCCG 59.8 0.23
mm4_intron_Gm872_chr10_92936953_F Primer3: not found at this Tm N.d. 0.23
mm4_intron_Gm872_chr10_92936953_R Primer3: not found at this Tm N.d. 0.23
mm4_intron_Igf1r_chr7_68160316_F TCGTCGGCAGCGTCTGCCCTTTAACCATGCCTGT 59.8 0.22
mm4_intron_Igf1r_chr7_68160316_R GTCTCGTGGGCTCGGCTGCTCTCCATGAGTGCCAA 60.0 0.22
mm4_intergenic_Tars|Npr3_chr15_11637578_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Tars|Npr3_chr15_11637578_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Rad51ap2|Gm23781_chr12_11649905_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Rad51ap2|Gm23781_chr12_11649905_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Mir446q|Tnik_chr3_28474332_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Mir446q|Tnik_chr3_28474332_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Kcnh8|Efhb_chr17_53042716_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Kcnh8|Efhb_chr17_53042716_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Gm7488|Tbl1xr1_chr3_21460757_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Gm7488|Tbl1xr1_chr3_21460757_R Primer3: not found at this Tm N.d. 0.22
mm4_exon_Acacb_chr5_114233116_F TCGTCGGCAGCGTCGTCTGGAAGAACCTGGAGGC 60.0 0.22
mm4_exon_Acacb_chr5_114233116_R GTCTCGTGGGCTCGGGGAAGGTGATGTCATTGCCG 59.2 0.22
mm4_intergenic_Nlgn2|Tmem256_chr11_69837855_F Primer3: not found at this Tm N.d. 0.21
mm4_intergenic_Nlgn2|Tmem256_chr11_69837855_R Primer3: not found at this Tm N.d. 0.21
mm4_intron_Plxna4_chr6_32227655_F TCGTCGGCAGCGTCCCCCAAGCAATAGGAGAGCC 60.1 0.21
mm4_intron_Plxna4_chr6_32227655_R GTCTCGTGGGCTCGGCTGCCTACCAGAACAGAACCA 59.6 0.21
mm4_exon_Nfatc1_chr18_80698420_F TCGTCGGCAGCGTCGTACGTCTTCCACCTCCACG 60.1 0.20
mm4_exon_Nfatc1_chr18_80698420_R GTCTCGTGGGCTCGGTTCCTGTCCTCTGGCAACAC 59.8 0.20
mm4_intergenic_Gm12439|Gm12438_chr4_48720101_F Primer3: not found at this Tm N.d. 0.20
mm4_intergenic_Gm12439|Gm12438_chr4_48720101_R Primer3: not found at this Tm N.d. 0.20
mm3_intergenic_Gm24280|Rbm26/Gm17066_chr14_105074328_F Primer3: not found at this Tm N.d. 0.20
mm3_intergenic_Gm24280|Rbm26/Gm17066_chr14_105074328_R Primer3: not found at this Tm N.d. 0.20
mm4_intergenic_Otol1|Gm23477_chr3_70052789_F Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Otol1|Gm23477_chr3_70052789_R Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Gm13503|Gm13502_chr2_53633264_F Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Gm13503|Gm13502_chr2_53633264_R Primer3: not found at this Tm N.d. 0.19
mm4_exon_Lamb3_chr1_193302013_F Primer3: not found at this Tm N.d. 0.18
mm4_exon_Lamb3_chr1_193302013_R Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Dact2|Smoc2_chr17_14222133_F TCGTCGGCAGCGTCTCCTAATCGCTAGGGGCTCA 59.8 0.18
mm4_intergenic_Dact2|Smoc2_chr17_14222133_R GTCTCGTGGGCTCGGTGAGTTGGTGGCATGAATGGA 59.9 0.18
mm4_intron_Adap1_chr5_139287251_F TCGTCGGCAGCGTCGTTCGTTCTGGGCAAGCATG 60.1 0.17
mm4_intron_Adap1_chr5_139287251_R GTCTCGTGGGCTCGGTGGGAAATGCAGAAGTCCCC 59.9 0.17
mm4_intergenic_Gm22183|mmu-mir-378d_chr10_126354871_F Primer3: not found at this Tm N.d. 0.17
mm4_intergenic_Gm22183|mmu-mir-378d_chr10_126354871_R Primer3: not found at this Tm N.d. 0.17
mm4_exon_Rftn2_chr1_55201884_F TCGTCGGCAGCGTCCACAGACTCGGACACTGGTC 60.0 0.17
mm4_exon_Rftn2_chr1_55201884_R GTCTCGTGGGCTCGGGCCAAACCTCTACTCACCCC 60.0 0.17
mm4_intergenic_Rbpms|Gm9951_chr8_33943147_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Rbpms|Gm9951_chr8_33943147_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm5793|4930468A15Rik_chrX_76560195_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm5793|4930468A15Rik_chrX_76560195_R Primer3: not found at this Tm N.d. 0.16
mm4_intron_Kifc3_chr8_95110662_F TCGTCGGCAGCGTCGAGACAGCCACATCCAGAGG 59.8 0.16
mm4_intron_Kifc3_chr8_95110662_R GTCTCGTGGGCTCGGCGACTGCGATCAGAACTGGT 60.1 0.16
mm4_intergenic_Pcdh20|Gm25886_chr14_88696093_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Pcdh20|Gm25886_chr14_88696093_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Maf/Gm15655|Dynlrb2_chr8_116497864_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Maf/Gm15655|Dynlrb2_chr8_116497864_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm4929|Fam84a_chr12_13917587_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm4929|Fam84a_chr12_13917587_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm25255|Csmd1_chr8_15641624_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm25255|Csmd1_chr8_15641624_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm23389|Gm23181_chr1_94921769_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm23389|Gm23181_chr1_94921769_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm23276|Dach1_chr14_97723586_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm23276|Dach1_chr14_97723586_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Cmtm6|Cmtm7_chr9_114750399_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Cmtm6|Cmtm7_chr9_114750399_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm2343|Kifap3_chr1_163740504_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm2343|Kifap3_chr1_163740504_R Primer3: not found at this Tm N.d. 0.15
mm4_intron_Gm12239_chr11_55797332_F TCGTCGGCAGCGTCTGCTGTAGGCAGGAGATCCTA 59.7 0.15
mm4_intron_Gm12239_chr11_55797332_R GTCTCGTGGGCTCGGGCTGGAGCTTCCTGTCTCAT 59.4 0.15
mm4_intron_Rnf40_chr7_127591871_F Primer3: not found at this Tm N.d. 0.14
mm4_intron_Rnf40_chr7_127591871_R Primer3: not found at this Tm N.d. 0.14
mm4_intergenic_Mir29b-1/Gm27019|Gm13834_chr6_31068370_F Primer3: not found at this Tm N.d. 0.14
mm4_intergenic_Mir29b-1/Gm27019|Gm13834_chr6_31068370_R Primer3: not found at this Tm N.d. 0.14
mm4_intergenic_Bmper|Npsr1_chr9_23812545_F Primer3: not found at this Tm N.d. 0.14
mm4_intergenic_Bmper|Npsr1_chr9_23812545_R Primer3: not found at this Tm N.d. 0.14
mm3_intergenic_Gm22788|Gm24432_chr18_34064848_F Primer3: not found at this Tm N.d. 0.14
mm3_intergenic_Gm22788|Gm24432_chr18_34064848_R Primer3: not found at this Tm N.d. 0.14
mm3_intergenic_Aldh1a3|Asb7_chr7_66626677_F Primer3: not found at this Tm N.d. 0.14
mm3_intergenic_Aldh1a3|Asb7_chr7_66626677_R Primer3: not found at this Tm N.d. 0.14
mm4_intron_Aqp8_chr7_123467310_F TCGTCGGCAGCGTCGACCAGAGAGGAAACTGCCC 60.0 0.13
mm4_intron_Aqp8_chr7_123467310_R GTCTCGTGGGCTCGGTTCCCCTGCAAGAAAGACCC 59.8 0.13
mm4_intron_Lama3_chr18_12359153_F TCGTCGGCAGCGTCCCACATCACCTTGGAGCTGT 59.9 0.13
mm4_intron_Lama3_chr18_12359153_R GTCTCGTGGGCTCGGAGCTGCCACAAGAAAGTCCA 59.8 0.13
mm3_intergenic_Gm13750|Gm25748_chr1_64174876_F Primer3: not found at this Tm N.d. 0.13
mm3_intergenic_Gm13750|Gm25748_chr1_64174876_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Padi1|4930515B02Rik_chr4_140871159_F TCGTCGGCAGCGTCCTCCAATGTCCTGGGAGCTC 59.8 0.13
mm4_intergenic_Padi1|4930515B02Rik_chr4_140871159_R GTCTCGTGGGCTCGGTGCCTCTGTGTTTCCGAGAC 59.9 0.13
mm4_intergenic_Abhd8|Mrpl34_chr8_71463790_F Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Abhd8|Mrpl34_chr8_71463790_R Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_9330159F19Rik|Echdc1_chr10_29243563_F Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_9330159F19Rik|Echdc1_chr10_29243563_R Primer3: not found at this Tm N.d. 0.12
mm4_intron_Tnfaip2_chr12_111443802_F TCGTCGGCAGCGTCGGAGATGGTGTGGGAAAGGG 60.0 0.12
mm4_intron_Tnfaip2_chr12_111443802_R GTCTCGTGGGCTCGGGTGATCCAGGAACAGCTCCC 60.1 0.12
mm4_intergenic_Suclg2|Fam19a1_chr6_96017858_F Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Suclg2|Fam19a1_chr6_96017858_R Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Tle3|Rplp1_chr9_61783371_F TCGTCGGCAGCGTCAGACACTCCAGGGTGCCTAT 59.9 0.12
mm4_intergenic_Tle3|Rplp1_chr9_61783371_R GTCTCGTGGGCTCGGTGGGACGAGAAGATGCCAAA 59.3 0.12
mm4_intergenic_Gm27025|Gm25700_chr13_86264331_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Gm27025|Gm25700_chr13_86264331_R Primer3: not found at this Tm N.d. 0.11
mm3_intergenic_Gm23829|Gm5974_chr1_48166818_F Primer3: not found at this Tm N.d. 0.10
mm3_intergenic_Gm23829|Gm5974_chr1_48166818_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_CT030233.1|Gm20675_chr14_67189162_F TCGTCGGCAGCGTCCGCCTAGGCTCTCTCTTCAC 59.6 0.10
mm4_intergenic_CT030233.1|Gm20675_chr14_67189162_R GTCTCGTGGGCTCGGGCCCAGGTTCAAATTGCCTT 59.3 0.10
mm2_intergenic_Isg20|Gm26633_chr7_78930718_F Primer3: not found at this Tm N.d. 0.10
mm2_intergenic_Isg20|Gm26633_chr7_78930718_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Vmn1r62|Vmn1r63_chr7_5788764_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Vmn1r62|Vmn1r63_chr7_5788764_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_VMN1R62|VMN1R63_chr7_GL456219_random_135298_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_VMN1R62|VMN1R63_chr7_GL456219_random_135298_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Spata19|Opcml_chr9_27684841_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Spata19|Opcml_chr9_27684841_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm22815|Cntn5_chr9_9403422_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm22815|Cntn5_chr9_9403422_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm14894|Gm5942_chrX_114679117_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm14894|Gm5942_chrX_114679117_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm13916|Gm13915_chr2_108489609_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm13916|Gm13915_chr2_108489609_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Fam173b|Tas2r119_chr15_31894006_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Fam173b|Tas2r119_chr15_31894006_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Luzp1_chr4_136504439_F Primer3: not found at this Tm N.d. 0.09
mm4_intron_Luzp1_chr4_136504439_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Fam163b_chr2_27118673_F TCGTCGGCAGCGTCCAGGGACCCTTCACATTCCC 60.0 0.09
mm4_intron_Fam163b_chr2_27118673_R GTCTCGTGGGCTCGGCGAGAGCAGAGCAGTCGATT 59.9 0.09
mm3_exon_Zbtb39/Gpr182_chr10_127742016_F TCGTCGGCAGCGTCCAGCCAGTGTGTCACTCTCA 59.6 0.09
mm3_exon_Zbtb39/Gpr182_chr10_127742016_R GTCTCGTGGGCTCGGTGCATCAGTAGGCAGCACAT 59.7 0.09
mm3_intergenic_Ttc27|Ltbp1_chr17_74989877_F Primer3: not found at this Tm N.d. 0.08
mm3_intergenic_Ttc27|Ltbp1_chr17_74989877_R Primer3: not found at this Tm N.d. 0.08
mm3_intergenic_Shcbp1|Gm9457_chr8_4808983_F Primer3: not found at this Tm N.d. 0.08
mm3_intergenic_Shcbp1|Gm9457_chr8_4808983_R Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_1700122O11Rik|1700067P10Rik_chr17_48038518_F TCGTCGGCAGCGTCCCATTTCCCCAGCTCCTCTG 60.1 0.08
mm4_intergenic_1700122O11Rik|1700067P10Rik_chr17_48038518_R GTCTCGTGGGCTCGGTCAGGCATTGCTGGTGCTAC 60.6 0.08
mm4_intron_Dgki_chr6_37150720_F TCGTCGGCAGCGTCCAGGCAACACATTAGCTCTGC 59.8 0.08
mm4_intron_Dgki_chr6_37150720_R GTCTCGTGGGCTCGGAGGAGCCCAAACTGAGGTTG 59.8 0.08
mm4_intron_Nhsl1_chr10_18392271_F TCGTCGGCAGCGTCATGAGAGCCAGCAGAGGAGA 60.0 0.08
mm4_intron_Nhsl1_chr10_18392271_R GTCTCGTGGGCTCGGCCTGCAACAGAAGTGGGAGT 59.8 0.08
mm4_intergenic_Gm25693|Gm12129_chr11_38059074_F Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Gm25693|Gm12129_chr11_38059074_R Primer3: not found at this Tm N.d. 0.08
mm4_exon_Nfrkb_chr9_31386253_F TCGTCGGCAGCGTCGACGAACCCCGGGTGAGA 60.9 0.07
mm4_exon_Nfrkb_chr9_31386253_R GTCTCGTGGGCTCGGTTCTCCCTTTCTCCCCGCTT 60.8 0.07
mm4_intron_Tdp1_chr12_99938015_F TCGTCGGCAGCGTCATCACTGCCAGCTCATGTCC 60.1 0.07
mm4_intron_Tdp1_chr12_99938015_R GTCTCGTGGGCTCGGAGAACGAGGACCACAGCTTG 59.9 0.07
mm4_intron_Tspan18_chr2_93256675_F TCGTCGGCAGCGTCTAGGACACCTAGCGCTTCCT 60.0 0.07
mm4_intron_Tspan18_chr2_93256675_R GTCTCGTGGGCTCGGCTCCTGTCCACCCAGTTGTC 59.9 0.07
mm4_intergenic_Gm13415/Pax8|Pax8_chr2_24443218_F TCGTCGGCAGCGTCTTTGAACATGGACCTGGCCA 59.8 0.07
mm4_intergenic_Gm13415/Pax8|Pax8_chr2_24443218_R GTCTCGTGGGCTCGGGGCCATGTCTCAGTTCCACA 59.9 0.07
mm4_intergenic_Asb1|Twist2_chr1_91755153_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Asb1|Twist2_chr1_91755153_R Primer3: not found at this Tm N.d. 0.06
mm3_intergenic_Mep1a|Ankrd66_chr17_43519025_F Primer3: not found at this Tm N.d. 0.05
mm3_intergenic_Mep1a|Ankrd66_chr17_43519025_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Rpl21-ps13|mmu-mir-6401_chr7_139736126_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Rpl21-ps13|mmu-mir-6401_chr7_139736126_R Primer3: not found at this Tm N.d. 0.05
mm4_intron_Zfp385b_chr2_77443079_F TCGTCGGCAGCGTCCCAAAACTGGCCTCACTGGA 60.1 0.05
mm4_intron_Zfp385b_chr2_77443079_R GTCTCGTGGGCTCGGAAGTGCCCAGGTGTAGTTGG 59.8 0.05
mm4_intron_Hal_chr10_93509723_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Hal_chr10_93509723_R Primer3: not found at this Tm N.d. 0.05
mm3_intergenic_Synj1|Synj1/4930404I05Rik_chr16_91010663_F Primer3: not found at this Tm N.d. 0.04
mm3_intergenic_Synj1|Synj1/4930404I05Rik_chr16_91010663_R Primer3: not found at this Tm N.d. 0.04
mm4_exon_Zfp800_chr6_28261723_F TCGTCGGCAGCGTCGCTCCACCTGCAACGGTC 61.0 0.04
mm4_exon_Zfp800_chr6_28261723_R GTCTCGTGGGCTCGGCACGAGAGAACGGGACTGAC 60.1 0.04
mm3_exon_Stab2_chr10_86841452_F Primer3: not found at this Tm N.d. 0.03
mm3_exon_Stab2_chr10_86841452_R Primer3: not found at this Tm N.d. 0.03
mm3_intergenic_Fndc3b|Tmem212_chr3_27782243_F TCGTCGGCAGCGTCAGCCAAGTACGGTGAACTGG 59.9 0.03
mm3_intergenic_Fndc3b|Tmem212_chr3_27782243_R GTCTCGTGGGCTCGGGGAGGGTAAGGGCTCTGTCA 60.6 0.03
mm4_intergenic_Gm8793|Nkain2_chr10_31769799_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm8793|Nkain2_chr10_31769799_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Sh3bp4_chr1_89137218_F TCGTCGGCAGCGTCGCTCTCTGTTCCTTGTGCCA 60.2 0.03
mm4_intron_Sh3bp4_chr1_89137218_R GTCTCGTGGGCTCGGCCTCAGACCGTGCAGTACAT 59.4 0.03
mm4_intron_Cdh13_chr8_119313457_F TCGTCGGCAGCGTCCTGTCACACGCAACCTCTCT 59.9 0.03
mm4_intron_Cdh13_chr8_119313457_R GTCTCGTGGGCTCGGTGCTGGGAAGGAAGTGTGTG 60.1 0.03
mm4_intron_Dgat2_chr7_99176681_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Dgat2_chr7_99176681_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Tmem163_chr1_127677900_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Tmem163_chr1_127677900_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Pdzd2|Gm24302_chr15_12638566_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Pdzd2|Gm24302_chr15_12638566_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_C030017K20Rik_chr4_155465946_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_C030017K20Rik_chr4_155465946_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Olig3|Ifngr1_chr10_19462145_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Olig3|Ifngr1_chr10_19462145_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Irx1|Rpl9-ps4_chr13_72341467_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Irx1|Rpl9-ps4_chr13_72341467_R Primer3: not found at this Tm N.d. 0.01
mm4_intron_Slco5a1_chr1_12948946_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Slco5a1_chr1_12948946_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Ncor2_chr5_125044178_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Ncor2_chr5_125044178_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Hivep3_chr4_120035909_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Hivep3_chr4_120035909_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Zfy2|Gm22119_chrY_2371505_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Zfy2|Gm22119_chrY_2371505_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Zfp335|Gm11458/Zfp335_chr2_164911238_F TCGTCGGCAGCGTCCGGGGTCAGTGGTTCACTC 60.0 0.00
mm4_intergenic_Zfp335|Gm11458/Zfp335_chr2_164911238_R GTCTCGTGGGCTCGGGACAGCCCCGAGACACCAG 62.3 0.00
mm4_intergenic_Gm22202|Tmem106c_chr15_97949184_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm22202|Tmem106c_chr15_97949184_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Asmt_chrX_170676326_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Asmt_chrX_170676326_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.

ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52313821 AGGGAGCCTGGGATTAGGAGCTTACTTGGGGGTTTTCCCCCTCCCCCCCTCTGGAGAGTCCGGGGATGGAGAGCCGAAAG
GACATG
mm4_intron_Pitpnb_chr5_111380806 AGTTGTGCCAGCACTTCAGTACAGAGACCTCCCCTCTGGAGAGTCGGTATCCTGACCACTTCTCTCAAGCA
mm4_intron_Ctnnd2_chr15_30302354 GAACGGACCTGCTTCACTGACCTCTGACCTGGAAGAGGAAGCTGGGAGCTGAACTGTCCCTTGCTCTGCTCAACACTCTC
CAGAGGGGTGG
ATGCTTTCTTGTTTCAGAACCGCT
mm4_intergenic_BC017158|Rgs10_chr7_128307285 TGGACATCTCTCCCACCTGGCCCCCTCTCTGGTGAGTCCTGGATATTTTTCTTTTCTCCAAGTTCTCTCTTCTCTGGAAC
ACCCCAAGGT
mm4_intron_Ppa1_chr10_61653066 CCATAAGACTCCGCCGTCTCAGAGAACCCTGAGAAATGATGTTTTGTAGGACAGCCAAGGGTCATCTCCATCCCCCCTTC
TGGAGTGTCCCGAG
GCAGCTGTGGTAACCCTTGGCTGAC
mm4_intergenic_Gm12066|Gm12068_chr11_24349134 GCGTATCAGAGGCTGAGTGTGATCCNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNGGCCCTGGGA
TCTTCCCCAGAGTCTCCAGAAAGGAGGCCACAGAGAGGGTGCCTGCTAATGACA
mm3_intergenic_Rora|F830001A07Rik_chr9_68823588 TATCGTGGTGGAGCTGATGCTTGAAAAACACATGTTTACCCAAAGTCTGGTGAGTTCCATGCCTCTGGAGAGTCCTGGGT
TTTGTTTCCTTGCGGGTCAGGCATGGTCAGTCAT
mm4_intron_Atg2b_chr12_105642191 ACCATTAGCTACCAACAAACCCAAATAAACATAAATATAGCTCAGGAGCAGACAGAGAACCAGGACCCTGCAGAGGGGAG
G
GGTTCCTTAAAACCCTGCTGG
mm4_intergenic_Syce1l|Adamts18_chr8_113688097 GATAGACAGGACAGAGCGGCAGCACATTCATCTTGCCTCAATCCTATTTCATGTCTTCCGGACTCTGTAGAGGGGCGGTA
GGTGGGAACAATTGATCTATTGAGAATTGCAGGCTGTATCTCCCTCT
mm3_intron_Plekha7_chr7_116288629 GGGATGACTACCTGGGGACTCAGTCCACCCCTCTGCTGAGTCCTGGGAGGAGGGACGGGCAGGAGGTAGTCCTCAGTAGT
GAGAGGAAGTTGCTGGC
mm4_intergenic_Peli2|Gm23881_chr14_48361056 TCTTTCCACGGTGTGCTGTTCACATGGGACTCTGTGTGGAAGACAATGGCCCATTCTTTTCTCTCAAGTTTAGCTTTCCA
GGAGGCCCCCTCTCTGGAGAGCGCTGGGTCCTTGTGGTGCCTGGGGTTCGAGAAATG
mm4_exon_Elavl4_chr4_110211633 GGCTTCTTCTGCCTCAATCCTCTTATCAAAGCGGATGAATCCCACCCCTCTAGAGACCCCTGGGGAAAGAAAAAGAACAT
TTGTAAAGAGTGAACAAATGTAATCACTGGGAAGTGCCCTGTGGATATTTTGCCT
mm4_intron_Zfp64_chr2_168911038 GGGGCAAGTAGCTGAAGATGACGGAGGAATGTGTAGACATATAAAGGTATCCCCCTTCCCCGGGCTCTTCAGAAGTGGGG
GATCTGCCAGTAGGAAGGCGGCACTCCACCTAAGGTAC
mm4_intron_Igf1r_chr7_68160316 TGCCCTTTAACCATGCCTGTGTGCCCCTCCTCTGGAGTGGCGGGGGAGGTCCCCTCCTTTCCCCCCATGGTGCTGTAATT
GGAAATTTGGTTACTGCACTAAATTTTAAGTTTTGGCACTCATGGAGAGCAG
mm4_exon_Acacb_chr5_114233116 GTCTGGAAGAACCTGGAGGCATTTCAATTAGCCTTCATCTCATTCCCAGGTGGGCATGGTGGCCTTCAAAATGAGGTTCA
AGACCCCGGAGTATCCAGAAGGCCGGGACGCCGTTGTCATCGGCAATGACATCACCTTCC
mm4_intron_Plxna4_chr6_32227655 CCCCAAGCAATAGGAGAGCCAAGTATGGAGGCATGCCTTCTGTCTGGCCTCTCCTCTTGAGAGTCCTGAGATGGTTCTGT
TCTGGTAGGCAG
mm4_exon_Nfatc1_chr18_80698420 GTACGTCTTCCACCTCCACGTCGTGGAAAAACTGGCCGCTGCCATGGTGTAGGCCCAGGTAGGAGGTGATCTCGATTCTC
GGACTCTCCAGAGTCGGGG
CCCCGTTGGGTCTGGTGTTGCCAGAGGACAGGAA
mm4_intergenic_Dact2|Smoc2_chr17_14222133 TCCTAATCGCTAGGGGCTCATGGTGCCCCGTGTACTCTCCAGAGAGGTGGTTTATTCTGTGACGATTACATTACACCTCA
ATAAGGCTGCTGGAAATTGAAAATGCAAACTGGGTTATCCATTCATGCCACCAACTCA
mm4_intron_Adap1_chr5_139287251 GTTCGTTCTGGGCAAGCATGGGGCCACTCCTCTGAAGAGTCCCGGCCTCACCTCTTTTGAGGCCCCACATTTCCCCTTTC
TCCATCTGAGGGGACTTCTGCATTTCCCA
mm4_exon_Rftn2_chr1_55201884 CACAGACTCGGACACTGGTCAGCCTCCCCTCTGCAGTGTCCTCGGAAAGTCCTAGTGACACACACTCCAGTGTCCAGACC
GCAGGCTGAGGGGGGTGAGTAGAGGTTTGGC
mm4_intron_Kifc3_chr8_95110662 GAGACAGCCACATCCAGAGGTGGAAGAGTGACCACGTCTTGACACTCANNNNNNNNNNNNNNNNNNNNNAGGACGGTCCT
GGG
GTCCTCACCAGTTCTGATCGCAGTCG
mm4_intron_Gm12239_chr11_55797332 TGCTGTAGGCAGGAGATCCTAGTGAACAAACACATTATTAATTGGGATGCTCCAAGCAATGCCCCCACTGAGGAGGAGTG
CCAGCCCTCTGGAGAGGTCAGGGGGCAGCATTGTTATGAGACAGGAAGCTCCAGC
mm4_intron_Aqp8_chr7_123467310 GACCAGAGAGGAAACTGCCCACCCACTGCCTGAGAGGACTCAGCTCAGGAAACCACCCGTCTAGAGAGTTCTGGGCTTAG
AGATTTGTAATGGAGAGCTGGGTCTTTCTTGCAGGGGAA
mm4_intron_Lama3_chr18_12359153 CCACATCACCTTGGAGCTGTGTCTATTCAAATCTCCTTCCATGTTAGCCCCGGAGTTTCCTGAGGGCGGGACTGTTTCTT
CCTGCTCCTGGACTTTCTTGTGGCAGCT
mm4_intergenic_Padi1|4930515B02Rik_chr4_140871159 CTCCAATGTCCTGGGAGCTCCTTGCGGGCTGAATCCCAACCTTGTAATTAAGTGTAGGGCTGGGCACTGCTTCCTGGACG
CTCCACAGGGGGGG
CAGTCTCGGAAACACAGAGGCA
mm4_intron_Tnfaip2_chr12_111443802 GGAGATGGTGTGGGAAAGGGCCCTTAAAGTGTGTTGTGTTTGGGCAAGCACCTCCCCTCCGCAGTTTCCGGGGTTCTCCT
TCGTGATGGTGGGTCTTCCCACTTGCCTTACCGGGAGCTGTTCCTGGATCAC
mm4_intergenic_Tle3|Rplp1_chr9_61783371 AGACACTCCAGGGTGCCTATGTTAGGTACAGCTATAAAGCACCCAGACCCCAGCTCTGGAGAGGCTGGGGGTGGGTGGAG
GGTGATGTCTGAATTTGGCATCTTCTCGTCCCA
mm4_intergenic_CT030233.1|Gm20675_chr14_67189162 CGCCTAGGCTCTCTCTTCACTTCCTGCCTCCTAACTCTCAAGAGGGGCGGAGCTGCTTTCTAGAAGGCAATTTGAACCTG
GGC
mm4_intron_Fam163b_chr2_27118673 CAGGGACCCTTCACATTCCCGGGCATGCTGCCTAACAGTTCTGCCCCCCTCGGCCTCTCCAGAGGCTGGGGCCAGCTGGC
CTTGCTAGGGTGAACCCAAACAGTCACTGGTGCCACTAGAATCGACTGCTCTGCTCTCG
mm3_exon_Zbtb39/Gpr182_chr10_127742016 CAGCCAGTGTGTCACTCTCAGTTGTCCCTCAGTAGATCCCACCCACCCCCTTGGAGAGTCCCGGGGGAGTGGGGAACACT
TCGGTCCTGATAGAAACTATGTGCTGCCTACTGATGCA
mm4_intergenic_1700122O11Rik|1700067P10Rik_chr17_48038518 CCATTTCCCCAGCTCCTCTGTGGAATGGATGAGATACCACAGAGCCCACAGTTCAGCTGGGGACCCCTCTCCTCTGAAGA
GTCAGGTG
CTGCTGGGATTAAAGGTAGCACCAGCAATGCCTGA
mm4_intron_Dgki_chr6_37150720 CAGGCAACACATTAGCTCTGCATAATTCCCCAGACTCATCAGACGGGTGGAATTTTTGAAAGAAGATGGTCCAGTCATGG
TTCCCTTTCCTCATGTTCTCCAAGATAGGTAGTTAACAACCTCAGTTTGGGCTCCT
mm4_intron_Nhsl1_chr10_18392271 ATGAGAGCCAGCAGAGGAGAGCAAAAGCGGAGAGGGTAGAGTGGGCCCAGTACTCTCCTCAGGGGAGGAGGGAGAATCAA
GTCAAGACTCCCACTTCTGTTGCAGG
mm4_exon_Nfrkb_chr9_31386253 GACGAACCCCGGGTGAGACCCGCGGACTCTGCTGAGGTGAGGGCGGGCCCAGGCGTCGGGTTTGGTGGGTGCCGGGGGAA
GGGGAGAAGCGGGGAGAAAGGGAGAA
mm4_intron_Tdp1_chr12_99938015 ATCACTGCCAGCTCATGTCCACGATCACCCAGGACCTCCCCTATGGAGGGCCCTGGGGAGGAACGAGAAACAAGCTGTGG
TCCTCGTTCT
mm4_intron_Tspan18_chr2_93256675 TAGGACACCTAGCGCTTCCTGGTCCGGATCATGAAGACTGTTCCATCAGTATCTCCAGAGTGGCCCAGGCTCTCGAGAGG
GGAGG
CAAAGGGCCACGGGACAACTGGGTGGACAGGAG
mm4_intergenic_Gm13415/Pax8|Pax8_chr2_24443218 TTTGAACATGGACCTGGCCAGAGCCTCTGTCCCCCCTGAGTCTCCTGAGGAGAGGTGTGGAACTGAGACATGGCC
mm4_intron_Zfp385b_chr2_77443079 CCAAAACTGGCCTCACTGGACACTGCCAAGGCTGACTCCTGGCACTCTCCAGATGGGAGGATCCAACTACACCTGGGCAC
TT
mm4_exon_Zfp800_chr6_28261723 GCTCCACCTGCAACGGTCCCCCGGGCTTTGGAGAGGGGCGGGGAGCCGCGCGGGTCCCCCACCTGGGGCCTTGCGGGTAG
CAGATGCTTCGGGAAACTTTATGGAAGTCAGTCCCGTTCTCTCGTG
mm3_intergenic_Fndc3b|Tmem212_chr3_27782243 AGCCAAGTACGGTGAACTGGGCCTCCCTTCAGTGTGGAAGTGCTTGTGACGCTGGCATAATGGATGCTGTATAAGAAGTC
TGCTGAGTTCTCCCCTCTGGAATCTCCGGGGCGCCTGACAGAGCCCTTACCCTCC
mm4_intron_Sh3bp4_chr1_89137218 GCTCTCTGTTCCTTGTGCCAAGCCTCCCACGGCCACTCCTCCAACAGGACCCGCCACTCTGGACAGACAGGGGCTGTTTT
CTTCCTGAGTCCTGGAAGTGCCAATGTACTGCACGGTCTGAGG
mm4_intron_Cdh13_chr8_119313457 CTGTCACACGCAACCTCTCTTTCCCCCTGCCTTTCCACAGGGGTGGGCAGAATTGAAATGTGGCCCACCACTGACCAGCG
ACCTTACTAATTACAAAGCACTTCAGAGTCCTGCTATCCCACACACTTCCTTCCCAGCA
mm4_intergenic_Zfp335|Gm11458/Zfp335_chr2_164911238 CGGGGTCAGTGGTTCACTCCCCCTCCCCGGACTCCCCTGCGGAGGGGGCTGACAGCGACGGTCGCGGGGCCTGGTGTCTC
GGGGCTGTC

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_mN1FkRligAThyqKv3eg3.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.