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Contents:

PCR primers for off-targets of CATCGACCGGCTCCTCCTCC TGG

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_52314035_F Primer3: not found at this Tm N.d. 1.00
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52314035_R Primer3: not found at this Tm N.d. 1.00
mm4_intron_Gm14014_chr2_106579731_F Primer3: not found at this Tm N.d. 0.41
mm4_intron_Gm14014_chr2_106579731_R Primer3: not found at this Tm N.d. 0.41
mm4_intron_Atp2b2_chr6_113903415_F TCGTCGGCAGCGTCCAGCTTGCTGGGCACATTAG 59.5 0.39
mm4_intron_Atp2b2_chr6_113903415_R GTCTCGTGGGCTCGGGGGTCTAAGCATGAGGGGTT 59.0 0.39
mm4_intron_Bbs2_chr8_94088843_F TCGTCGGCAGCGTCTCCGAAGCATCAGAAGTGGT 59.0 0.36
mm4_intron_Bbs2_chr8_94088843_R GTCTCGTGGGCTCGGAGTGCTCACTGTGTGCATCT 59.6 0.36
mm4_intron_Pawr_chr10_108404877_F Primer3: not found at this Tm N.d. 0.36
mm4_intron_Pawr_chr10_108404877_R Primer3: not found at this Tm N.d. 0.36
mm3_intergenic_Unc5d|4933433F19Rik_chr8_29277711_F Primer3: not found at this Tm N.d. 0.32
mm3_intergenic_Unc5d|4933433F19Rik_chr8_29277711_R Primer3: not found at this Tm N.d. 0.32
mm4_intergenic_Gm26005|Epcam_chr17_87508122_F TCGTCGGCAGCGTCACCTGCTGCGAATAGACCTG 59.8 0.32
mm4_intergenic_Gm26005|Epcam_chr17_87508122_R GTCTCGTGGGCTCGGGAGGGTTTGCTAAGTGGGCT 59.9 0.32
mm4_intron_Vmn2r91_chr17_18110892_F Primer3: not found at this Tm N.d. 0.31
mm4_intron_Vmn2r91_chr17_18110892_R Primer3: not found at this Tm N.d. 0.31
mm4_intron_Phf15_chr11_51855632_F Primer3: not found at this Tm N.d. 0.31
mm4_intron_Phf15_chr11_51855632_R Primer3: not found at this Tm N.d. 0.31
mm4_intron_Cldn15_chr5_136970859_F TCGTCGGCAGCGTCCCCAAGCCTTTTGCCTTCTG 59.6 0.31
mm4_intron_Cldn15_chr5_136970859_R GTCTCGTGGGCTCGGCCTTTCCCCAAAGCTGAGGG 60.6 0.31
mm4_intron_Gm14486_chr2_30660836_F Primer3: not found at this Tm N.d. 0.30
mm4_intron_Gm14486_chr2_30660836_R Primer3: not found at this Tm N.d. 0.30
mm3_intergenic_Crispld1|Crisp4_chr1_18058811_F Primer3: not found at this Tm N.d. 0.30
mm3_intergenic_Crispld1|Crisp4_chr1_18058811_R Primer3: not found at this Tm N.d. 0.30
mm4_exon_Megf8_chr7_25346120_F TCGTCGGCAGCGTCCTCTGCCCACCTCTCTACCT 60.0 0.29
mm4_exon_Megf8_chr7_25346120_R GTCTCGTGGGCTCGGAGGACCTCCTGATCCACACC 60.6 0.29
mm4_intergenic_5430402E10Rik|Gm14750_chrX_77950626_F Primer3: not found at this Tm N.d. 0.29
mm4_intergenic_5430402E10Rik|Gm14750_chrX_77950626_R Primer3: not found at this Tm N.d. 0.29
mm4_intergenic_Gxylt2|Gm15576_chr6_100711362_F Primer3: not found at this Tm N.d. 0.29
mm4_intergenic_Gxylt2|Gm15576_chr6_100711362_R Primer3: not found at this Tm N.d. 0.29
mm4_intron_Kcnh6_chr11_106012365_F TCGTCGGCAGCGTCGGGGTAATCTAGGGGCCAGA 60.1 0.27
mm4_intron_Kcnh6_chr11_106012365_R GTCTCGTGGGCTCGGTCGAGGAATGGCAAAGTTCCA 59.9 0.27
mm4_intergenic_4930569F06Rik|Suds3_chr5_116921074_F TCGTCGGCAGCGTCCATTACCCAGCACATTCCCC 58.5 0.26
mm4_intergenic_4930569F06Rik|Suds3_chr5_116921074_R GTCTCGTGGGCTCGGACTTCGAAGGACAGAGTGGA 58.0 0.26
mm4_exon_Myo18b_chr5_112785579_F TCGTCGGCAGCGTCCTGTGGCCAGAGCTTACACA 59.9 0.26
mm4_exon_Myo18b_chr5_112785579_R GTCTCGTGGGCTCGGCCAGCGGTTTGAGTTGGAGA 60.2 0.26
mm4_intergenic_Gm17033|4933406K04Rik_chr12_106684685_F Primer3: not found at this Tm N.d. 0.26
mm4_intergenic_Gm17033|4933406K04Rik_chr12_106684685_R Primer3: not found at this Tm N.d. 0.26
mm4_intron_Meig1_chr2_3409328_F Primer3: not found at this Tm N.d. 0.25
mm4_intron_Meig1_chr2_3409328_R Primer3: not found at this Tm N.d. 0.25
mm4_intergenic_Rassf6|Cxcl5_chr5_90659358_F Primer3: not found at this Tm N.d. 0.25
mm4_intergenic_Rassf6|Cxcl5_chr5_90659358_R Primer3: not found at this Tm N.d. 0.25
mm4_intergenic_Sfrp1|Zmat4_chr8_23463720_F TCGTCGGCAGCGTCGAGCAGCTCCCCTTCACAAG 60.6 0.25
mm4_intergenic_Sfrp1|Zmat4_chr8_23463720_R GTCTCGTGGGCTCGGTCCTCTTCTCCAGGTCAGGG 59.9 0.25
mm4_intergenic_Rhpn1|Mafa_chr15_75732671_F TCGTCGGCAGCGTCGGTTGTGGGCTGGATGAGAA 59.9 0.22
mm4_intergenic_Rhpn1|Mafa_chr15_75732671_R GTCTCGTGGGCTCGGTCATCAGGTCCACTTGTGCC 59.9 0.22
mm4_intergenic_Gm15647|Bicc1_chr10_71106955_F Primer3: not found at this Tm N.d. 0.21
mm4_intergenic_Gm15647|Bicc1_chr10_71106955_R Primer3: not found at this Tm N.d. 0.21
mm3_exon_Rhbdf2_chr11_116604412_F Primer3: not found at this Tm N.d. 0.19
mm3_exon_Rhbdf2_chr11_116604412_R Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Chrna10|Nup98_chr7_102121773_F TCGTCGGCAGCGTCCAGTGACATTAAAGACAAACCTGA 57.3 0.19
mm4_intergenic_Chrna10|Nup98_chr7_102121773_R GTCTCGTGGGCTCGGTCCTTTGATGCGTCCTTCCA 59.3 0.19
mm4_intron_Spata13_chr14_60634387_F TCGTCGGCAGCGTCCTTGGAGTGAGTGAGTGGCA 59.6 0.19
mm4_intron_Spata13_chr14_60634387_R GTCTCGTGGGCTCGGTCATTCCCCGCCTCTCACTA 60.0 0.19
mm4_exon_Fxr2_chr11_69633212_F Primer3: not found at this Tm N.d. 0.18
mm4_exon_Fxr2_chr11_69633212_R Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Pfas|Ctc1_chr11_69012461_F Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Pfas|Ctc1_chr11_69012461_R Primer3: not found at this Tm N.d. 0.18
mm4_exon_Mdn1_chr4_32760824_F TCGTCGGCAGCGTCATCCAGCCATGCAGAAGAGC 60.4 0.17
mm4_exon_Mdn1_chr4_32760824_R GTCTCGTGGGCTCGGGGGCTGAAGACCTTGGTCAA 59.8 0.17
mm4_exon_Ezh1_chr11_101192684_F TCGTCGGCAGCGTCGCTTGGCAGCTGAGGAAAAC 60.0 0.17
mm4_exon_Ezh1_chr11_101192684_R GTCTCGTGGGCTCGGGGAAACAGAGGCGGTGAAGA 59.9 0.17
mm4_intron_Pla2g6_chr15_79304980_F Primer3: not found at this Tm N.d. 0.17
mm4_intron_Pla2g6_chr15_79304980_R Primer3: not found at this Tm N.d. 0.17
mm4_intergenic_Gm12490|Cd2_chr3_101272525_F Primer3: not found at this Tm N.d. 0.17
mm4_intergenic_Gm12490|Cd2_chr3_101272525_R Primer3: not found at this Tm N.d. 0.17
mm4_exon_Dok5_chr2_170731824_F Primer3: not found at this Tm N.d. 0.16
mm4_exon_Dok5_chr2_170731824_R Primer3: not found at this Tm N.d. 0.16
mm4_intron_Hmcn1_chr1_150979786_F TCGTCGGCAGCGTCAGAAGAGCAAGAGAAGCGGC 60.3 0.16
mm4_intron_Hmcn1_chr1_150979786_R GTCTCGTGGGCTCGGAAGGGCAGATTTCCAGCTCC 60.0 0.16
mm4_intergenic_Gm807|Zfp366_chr13_99159150_F TCGTCGGCAGCGTCACAGCCTTCAGAGACACTGC 59.9 0.16
mm4_intergenic_Gm807|Zfp366_chr13_99159150_R GTCTCGTGGGCTCGGGCCTTTGGGAGTGAGCAATG 59.4 0.16
mm4_intron_Mybpc1_chr10_88591780_F Primer3: not found at this Tm N.d. 0.15
mm4_intron_Mybpc1_chr10_88591780_R Primer3: not found at this Tm N.d. 0.15
mm3_intergenic_Zdhhc16|Mms19/Zdhhc16_chr19_41940762_F Primer3: not found at this Tm N.d. 0.15
mm3_intergenic_Zdhhc16|Mms19/Zdhhc16_chr19_41940762_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Zc3h13|Siah3_chr14_75433050_F TCGTCGGCAGCGTCTTAGGACAGTCACTGGCAGC 59.6 0.13
mm4_intergenic_Zc3h13|Siah3_chr14_75433050_R GTCTCGTGGGCTCGGCACCACGGGCATTTTCCTTC 59.7 0.13
mm4_intergenic_Ifnl3|Sycn_chr7_28527231_F Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Ifnl3|Sycn_chr7_28527231_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Csrnp1|Xirp1_chr9_119984668_F TCGTCGGCAGCGTCTCTCTCCCGTTCTGACTGCA 60.5 0.12
mm4_intergenic_Csrnp1|Xirp1_chr9_119984668_R GTCTCGTGGGCTCGGGGAGGGTCGAGCTTCCAC 59.4 0.12
mm4_exon_Olfr380_chr11_73454164_F TCGTCGGCAGCGTCGGACAGGAACAGGGCAAAGA 59.8 0.12
mm4_exon_Olfr380_chr11_73454164_R GTCTCGTGGGCTCGGTGGAGAGGAAGAATGATAAAGAACA 57.7 0.12
mm4_exon_Ubr5_chr15_38078517_F Primer3: not found at this Tm N.d. 0.12
mm4_exon_Ubr5_chr15_38078517_R Primer3: not found at this Tm N.d. 0.12
mm4_exon_Kctd17_chr15_78437247_F Primer3: not found at this Tm N.d. 0.12
mm4_exon_Kctd17_chr15_78437247_R Primer3: not found at this Tm N.d. 0.12
mm4_intron_Vsig1_chrX_140918271_F TCGTCGGCAGCGTCTGCCCATGCTGAGGAAGAAC 60.3 0.12
mm4_intron_Vsig1_chrX_140918271_R GTCTCGTGGGCTCGGGGGCTGTGATGGGGATATGG 59.9 0.12
mm4_intergenic_Gm12991|Ncmap_chr4_135379158_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Gm12991|Ncmap_chr4_135379158_R Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Txn1|Txndc8_chr4_57967340_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Txn1|Txndc8_chr4_57967340_R Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Gm11367|Mboat1_chr13_30136442_F TCGTCGGCAGCGTCGTCTCCCGGTCTCCTGCC 61.4 0.11
mm4_intergenic_Gm11367|Mboat1_chr13_30136442_R GTCTCGTGGGCTCGGACGTTCTGCTTCCCGCTC 60.0 0.11
mm3_intron_Thsd7b_chr1_130091351_F Primer3: not found at this Tm N.d. 0.10
mm3_intron_Thsd7b_chr1_130091351_R Primer3: not found at this Tm N.d. 0.10
mm3_intron_Tnni3k_chr3_154811496_F TCGTCGGCAGCGTCTGATGCGGCCTTAGTGTGTT 59.9 0.10
mm3_intron_Tnni3k_chr3_154811496_R GTCTCGTGGGCTCGGTGACAGGCGTGAGATCACAG 59.7 0.10
mm4_intergenic_Glt1d1|Tmem132d_chr5_127756868_F TCGTCGGCAGCGTCAACAAATGTGGTGCCATGGC 59.9 0.10
mm4_intergenic_Glt1d1|Tmem132d_chr5_127756868_R GTCTCGTGGGCTCGGTCTGGACCCATCTGCACATG 59.7 0.10
mm4_exon_Fam126a_chr5_23964406_F TCGTCGGCAGCGTCGTAGCATCCCCACTGCTCTC 59.8 0.10
mm4_exon_Fam126a_chr5_23964406_R GTCTCGTGGGCTCGGTGTGAGAGGACTGGGCTGTA 59.8 0.10
mm4_intergenic_1700017N19Rik|Gm26278_chr10_101524005_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_1700017N19Rik|Gm26278_chr10_101524005_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Tjp2|Fxn_chr19_24249566_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Tjp2|Fxn_chr19_24249566_R Primer3: not found at this Tm N.d. 0.09
mm4_exon_Foxd1_chr13_98354822_F Primer3: not found at this Tm N.d. 0.09
mm4_exon_Foxd1_chr13_98354822_R Primer3: not found at this Tm N.d. 0.09
mm3_intergenic_Insig1|En2_chr5_28138257_F TCGTCGGCAGCGTCTGCATGACTTTTCCCCACCT 59.5 0.09
mm3_intergenic_Insig1|En2_chr5_28138257_R GTCTCGTGGGCTCGGGGCCATGGTTGTATCCACCA 60.0 0.09
mm4_intergenic_Gm1862|Gm14591_chrX_53948056_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm1862|Gm14591_chrX_53948056_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_RP24-86O15.2|Gm10698_chr9_33426252_F TCGTCGGCAGCGTCGCATCGTACTGGGTTGTGGA 60.0 0.09
mm4_intergenic_RP24-86O15.2|Gm10698_chr9_33426252_R GTCTCGTGGGCTCGGTCCAATTGCTGAAGTGGGGA 59.2 0.09
mm4_intergenic_Gm23225|Gm24127_chr14_91619118_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm23225|Gm24127_chr14_91619118_R Primer3: not found at this Tm N.d. 0.09
mm3_intron_Anks1b_chr10_90832864_F Primer3: not found at this Tm N.d. 0.09
mm3_intron_Anks1b_chr10_90832864_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Mypn_chr10_63122526_F Primer3: not found at this Tm N.d. 0.09
mm4_intron_Mypn_chr10_63122526_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_2610307P16Rik|mmu-mir-6368_chr13_28698100_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_2610307P16Rik|mmu-mir-6368_chr13_28698100_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm25261|Gm831_chr4_131428225_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm25261|Gm831_chr4_131428225_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_6530411M01Rik|Gm8492_chr17_9173834_F TCGTCGGCAGCGTCCCTCAGACTGTTCCGAAGCA 59.6 0.08
mm4_intergenic_6530411M01Rik|Gm8492_chr17_9173834_R GTCTCGTGGGCTCGGAGCCGTTTCTCAGACCAGTT 59.2 0.08
mm4_intergenic_Mapk10|Ptpn13_chr5_103352208_F Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Mapk10|Ptpn13_chr5_103352208_R Primer3: not found at this Tm N.d. 0.08
mm3_exon_Incenp_chr19_9885741_F TCGTCGGCAGCGTCTACCCATCTCTGGCCTGTGA 59.9 0.07
mm3_exon_Incenp_chr19_9885741_R GTCTCGTGGGCTCGGGTTCTCACAGTGCTCCTGCT 59.9 0.07
mm4_intron_Prepl_chr17_85089247_F TCGTCGGCAGCGTCACTGATTCAGTGAACCGCCC 60.3 0.07
mm4_intron_Prepl_chr17_85089247_R GTCTCGTGGGCTCGGTGGTGAAATCCAATTGCTCCT 57.8 0.07
mm4_intergenic_Rap1a|Adora3_chr3_105806971_F TCGTCGGCAGCGTCAATCCTACCCAGGCCATTTAC 57.4 0.07
mm4_intergenic_Rap1a|Adora3_chr3_105806971_R GTCTCGTGGGCTCGGGTCGGTAGATACGGCGTCTG 60.0 0.07
mm4_intergenic_Npm1|Gm12115_chr11_33163227_F TCGTCGGCAGCGTCGCCTTTCATCCTCACGCCTA 59.8 0.07
mm4_intergenic_Npm1|Gm12115_chr11_33163227_R GTCTCGTGGGCTCGGCTTGGGCGACAGTAGTGGAG 60.1 0.07
mm4_intergenic_Gm25460|Gm5263_chr1_145842508_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Gm25460|Gm5263_chr1_145842508_R Primer3: not found at this Tm N.d. 0.07
mm4_intron_Zfp532_chr18_65656662_F Primer3: not found at this Tm N.d. 0.07
mm4_intron_Zfp532_chr18_65656662_R Primer3: not found at this Tm N.d. 0.07
mm4_exon_4833427F10Rik_chr17_35776866_F Primer3: not found at this Tm N.d. 0.07
mm4_exon_4833427F10Rik_chr17_35776866_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_n-R5s79|Gm4925/Arl1_chr10_88711494_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_n-R5s79|Gm4925/Arl1_chr10_88711494_R Primer3: not found at this Tm N.d. 0.06
mm3_intergenic_Ttc39c|Mir1948_chr18_12704592_F Primer3: not found at this Tm N.d. 0.06
mm3_intergenic_Ttc39c|Mir1948_chr18_12704592_R Primer3: not found at this Tm N.d. 0.06
mm4_exon_Espnl_chr1_91327564_F TCGTCGGCAGCGTCTTCACAGACATCGGCCTGAC 60.0 0.06
mm4_exon_Espnl_chr1_91327564_R GTCTCGTGGGCTCGGCCCTCCCCAAGAGTCTCTCA 59.9 0.06
mm4_intergenic_Gm6027|Gm4746_chrX_90326945_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm6027|Gm4746_chrX_90326945_R Primer3: not found at this Tm N.d. 0.06
mm4_intron_Lphn3_chr5_81062282_F TCGTCGGCAGCGTCCAGCTTGTGTCCTGTTGCAG 59.6 0.05
mm4_intron_Lphn3_chr5_81062282_R GTCTCGTGGGCTCGGGAACCCCTGAGAACTGTGGG 59.9 0.05
mm4_intergenic_Gm13375|Gm13377_chr2_21028402_F TCGTCGGCAGCGTCTCCTGCTGGAGGAGCTGAAA 60.8 0.05
mm4_intergenic_Gm13375|Gm13377_chr2_21028402_R GTCTCGTGGGCTCGGCTGGGGTCATGTGACATCCC 60.1 0.05
mm4_intergenic_Vps13c|Gm22382_chr9_68126396_F TCGTCGGCAGCGTCAGCCAAGAGTGAATCATGCTGA 60.0 0.05
mm4_intergenic_Vps13c|Gm22382_chr9_68126396_R GTCTCGTGGGCTCGGGCCCTTCTTGCTTCCCAGAA 60.2 0.05
mm4_exon_Rpusd3_chr6_113415625_F TCGTCGGCAGCGTCTGCAGGGTCCGAGAGAAGTA 59.9 0.05
mm4_exon_Rpusd3_chr6_113415625_R GTCTCGTGGGCTCGGCCGCCTAAGTCCTTCCCAAG 60.1 0.05
mm4_intergenic_Clec14a|Gm17529_chr12_58801268_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Clec14a|Gm17529_chr12_58801268_R Primer3: not found at this Tm N.d. 0.04
mm3_exon_Tmc5_chr7_118656784_F Primer3: not found at this Tm N.d. 0.04
mm3_exon_Tmc5_chr7_118656784_R Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Gm13266|Gm10849_chr2_14557243_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Gm13266|Gm10849_chr2_14557243_R Primer3: not found at this Tm N.d. 0.04
mm3_intron_A830082K12Rik_chr13_78250814_F TCGTCGGCAGCGTCTGACCCATGATGGTGTCTCT 58.0 0.04
mm3_intron_A830082K12Rik_chr13_78250814_R GTCTCGTGGGCTCGGAAAGGTGTCCAATGGCCTCA 59.5 0.04
mm4_exon_Spata22/Olfr20_chr11_73353777_F TCGTCGGCAGCGTCCCAGCATGGCAGAAGGGAA 60.0 0.04
mm4_exon_Spata22/Olfr20_chr11_73353777_R GTCTCGTGGGCTCGGGGACGGTGGTGAGGTACATG 60.1 0.04
mm4_exon_Fam20a_chr11_109722142_F TCGTCGGCAGCGTCGCGTCTCTTCCAAGGATGCT 60.1 0.04
mm4_exon_Fam20a_chr11_109722142_R GTCTCGTGGGCTCGGTGTCAACTCTGCGTTCCCTC 59.9 0.04
mm4_intron_Zfp618_chr4_63068699_F Primer3: not found at this Tm N.d. 0.04
mm4_intron_Zfp618_chr4_63068699_R Primer3: not found at this Tm N.d. 0.04
mm4_intron_Gmpr_chr13_45530846_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Gmpr_chr13_45530846_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Ifltd1_chr6_145613409_F TCGTCGGCAGCGTCGTCCAAGGAGAGACCGGTTG 60.0 0.03
mm4_intron_Ifltd1_chr6_145613409_R GTCTCGTGGGCTCGGAGAATGCCACCCACCTCATC 59.7 0.03
mm3_intron_Adam28_chr14_68632383_F Primer3: not found at this Tm N.d. 0.03
mm3_intron_Adam28_chr14_68632383_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm9581|Arl6_chr16_59601134_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm9581|Arl6_chr16_59601134_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_9130401M01Rik/Zhx1_chr15_58076377_F TCGTCGGCAGCGTCGAGAGACTCCGCTCCCTGAA 60.6 0.02
mm4_exon_9130401M01Rik/Zhx1_chr15_58076377_R GTCTCGTGGGCTCGGGGAGTCCTCGAGTGGGGT 59.9 0.02
mm4_exon_Mrm1_chr11_84819152_F TCGTCGGCAGCGTCCTGCAGCCCAGCCTTACC 60.4 0.02
mm4_exon_Mrm1_chr11_84819152_R GTCTCGTGGGCTCGGAGCGGCTGGAACGTCTATTT 59.7 0.02
mm4_intergenic_Fam76b|Sesn3_chr9_13945093_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Fam76b|Sesn3_chr9_13945093_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Ggt5_chr10_75609200_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Ggt5_chr10_75609200_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Olfr376_chr11_73374782_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Olfr376_chr11_73374782_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Olfr1_chr11_73395974_F TCGTCGGCAGCGTCGGACGGTGGTGAGGTACATG 60.1 0.02
mm4_exon_Olfr1_chr11_73395974_R GTCTCGTGGGCTCGGAGACACTATGACAGAAAGGAACA 57.1 0.02
mm4_intron_Fam20a_chr11_109687493_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Fam20a_chr11_109687493_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Slc45a1_chr4_150636752_F TCGTCGGCAGCGTCTGCTCCTAGAAACAGCAGGC 60.0 0.02
mm4_intron_Slc45a1_chr4_150636752_R GTCTCGTGGGCTCGGCAGTGAGACACTCTGCCCAG 60.0 0.02
mm4_exon_Ube2r2_chr4_41136125_F TCGTCGGCAGCGTCCAGATGACCAGCTCGCAGAA 60.1 0.02
mm4_exon_Ube2r2_chr4_41136125_R GTCTCGTGGGCTCGGGCCACCTCCCAGTTGTAGAG 59.7 0.02
mm4_intergenic_Phlpp1|Bcl2/D630008O14Rik_chr1_106400614_F TCGTCGGCAGCGTCCATTTGAGCTGGGCTTGCTG 60.1 0.02
mm4_intergenic_Phlpp1|Bcl2/D630008O14Rik_chr1_106400614_R GTCTCGTGGGCTCGGTCTGAGGGTACTGGGGACTG 59.9 0.02
mm3_intergenic_Zswim6|Gm22118_chr13_107793645_F Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_Zswim6|Gm22118_chr13_107793645_R Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_Atxn10|Gm23517_chr15_85350981_F Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_Atxn10|Gm23517_chr15_85350981_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Pias1_chr9_62886113_F TCGTCGGCAGCGTCCCAGATTGAAGCCCAAGGGT 59.9 0.01
mm4_intron_Pias1_chr9_62886113_R GTCTCGTGGGCTCGGTGTGATGCCACCAGACACTT 59.5 0.01
mm4_exon_Adrbk1_chr19_4305983_F Primer3: not found at this Tm N.d. 0.01
mm4_exon_Adrbk1_chr19_4305983_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Syne2|Esr2_chr12_76111247_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Syne2|Esr2_chr12_76111247_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Olfr23_chr11_73940270_F TCGTCGGCAGCGTCTGCCTGGAAAGAACCAAACTG 58.9 0.01
mm4_exon_Olfr23_chr11_73940270_R GTCTCGTGGGCTCGGGGCATAGAACAGGTGCTGGT 60.0 0.01
mm4_exon_Frrs1_chr3_116899140_F TCGTCGGCAGCGTCGGCCGTCTTTGCACCATTTT 59.9 0.01
mm4_exon_Frrs1_chr3_116899140_R GTCTCGTGGGCTCGGGGGCAAAACAAAAGCAACGC 59.9 0.01
mm4_intergenic_Gm24274|Gm24677_chr8_67277858_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm24274|Gm24677_chr8_67277858_R Primer3: not found at this Tm N.d. 0.01
mm4_intron_Aqp3_chr4_41096442_F TCGTCGGCAGCGTCCGGGCACTATGTCCAGTCAG 60.1 0.01
mm4_intron_Aqp3_chr4_41096442_R GTCTCGTGGGCTCGGGATTAGCCCCAAGGTGGTGG 60.3 0.01
mm4_exon_Nt5m_chr11_59848109_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Nt5m_chr11_59848109_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm23630|Mir129-2_chr2_94233035_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm23630|Mir129-2_chr2_94233035_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Oxct1_chr15_4135230_F TCGTCGGCAGCGTCTTCGGCCTTTACAGAGAGCG 60.1 0.00
mm4_intron_Oxct1_chr15_4135230_R GTCTCGTGGGCTCGGCTCAAGGGAAGAGCAGCCAA 59.9 0.00
mm3_intergenic_Foxn4|Myo1h_chr5_114295047_F Primer3: not found at this Tm N.d. 0.00
mm3_intergenic_Foxn4|Myo1h_chr5_114295047_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_intron_Atp2b2_chr6_113903415 CAGCTTGCTGGGCACATTAGCTTCCATTTTCAGATGAGCTTCCTGGAGGAGGAGCTGGTTGTTTGTGTAACGCAATGACG
TTCTTGTAATCACTATCCTCAAAGAGGAGGGTCTAACCCCTCATGCTTAGACCC
mm4_intron_Bbs2_chr8_94088843 TCCGAAGCATCAGAAGTGGTTTATAAGATTACAGATGCGACTGAGGTCCCGCCAGCCTTGGCCGTCTCCTCCTCCAGGGT
CTCAGAGATGCACACAGTGAGCACT
mm4_intergenic_Gm26005|Epcam_chr17_87508122 ACCTGCTGCGAATAGACCTGGAGGACTGGTTGTTAAGGGAACACCTCCTGGAACACTCACTTGCTCCCTCCATCAGCTCC
TCCTCCCGG
CCTGCCTCCCAGGGCCTGGGCTGTAACACAGCCCACTTAGCAAACCCTC
mm4_intron_Cldn15_chr5_136970859 CCCAAGCCTTTTGCCTTCTGCACCATGCACCATGCACTATGGGCCCCACCCAGAGAAGGGGTACAAGGTTCCTGAGAGTG
TTTGGCAACCATCAAAACCAGGAGGAGGAGCCAGACGCTTCCCTCAGCTTTGGGGAAAGG
mm4_exon_Megf8_chr7_25346120 CTCTGCCCACCTCTCTACCTTTCTCCACAGGGTTACTTGTGTTGCACTGGGAAGCCAACGGCTCCTCCTCCTGGGGATTC
AATGCTTCTGTGGGCTCTGCCCGGTGTGGATCAGGAGGTCCT
mm4_intron_Kcnh6_chr11_106012365 GGGGTAATCTAGGGGCCAGACAACTCCTCTTAATCAGAGGAGTTCGTGGTGAGAGGAGCGTCTTGGAGGAAGAATCAGGC
AGAGACTGGCTCTTCCTCCAGG
GACAGAGCAAAGACACTGGAACTTTGCCATTCCTCGA
mm4_intergenic_4930569F06Rik|Suds3_chr5_116921074 CATTACCCAGCACATTCCCCCATTACCTGGCTCCTCCCCCTTGCCCGACTCCTCCTCCTGGCACCTCCCCTTTTACTCTT
TTACTTGCTACTCCTCAATCCACTCTGTCCTTCGAAGT
mm4_exon_Myo18b_chr5_112785579 CTGTGGCCAGAGCTTACACACCCGCTTCTGGCAGGACTGACGCACATCCTCCAGCTCTTCCTCCTGGTCCTCACGGTCTT
TCTGATGCATCTGCTTCATCCTCTCAATCTCCAACTCAAACCGCTGG
mm4_intergenic_Sfrp1|Zmat4_chr8_23463720 GAGCAGCTCCCCTTCACAAGGTACCTTTCTCAGGAGAGAGTCGAGGACTGGCTCCTCCTCCGGGATGTTAGTGGAGCTGC
CCTGACCTGGAGAAGAGGA
mm4_intergenic_Rhpn1|Mafa_chr15_75732671 GGTTGTGGGCTGGATGAGAACAGAGGAAAGGCTGTAAGACAACCCCAACCCCTGGCTCCTCCTCCAGGCATTCCTGTGGC
TACCTGTGGGAGCCCAGAGAGGCACAAGTGGACCTGATGA
mm4_intergenic_Chrna10|Nup98_chr7_102121773 CAGTGACATTAAAGACAAACCTGATTTACTTTTCTCCCTCCACCGGCTCTTCCTACAGGCTACTCAGTGTCTGAATGCAT
GCCATGGAAGGACGCATCAAAGGA
mm4_intron_Spata13_chr14_60634387 CTTGGAGTGAGTGAGTGGCACGAAGGCATAACCTGGAGGTGGAGCGGGTGGAGGTAGTGAGAGGCGGGGAATGA
mm4_exon_Mdn1_chr4_32760824 ATCCAGCCATGCAGAAGAGCATTCCCTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCAAGG
CCACCACTGATGGAGGTGGAGAAAGTGGTGTCAACCCTGTTGACCAAGGTCTTCAGCCC
mm4_exon_Ezh1_chr11_101192684 GCTTGGCAGCTGAGGAAAACTCAAGCCCTGGCATTATCCCACACCCACCATCTCCTCCTCCTGGCATGGGAGCGCAGCTT
CTTACATCTGAACTCTCCTGGGAGACCAGGTCTTCACCGCCTCTGTTTCC
mm4_intron_Hmcn1_chr1_150979786 AGAAGAGCAAGAGAAGCGGCCCTGGAGGAGGAGCTGGCCCTTGCGAGTCAGAAACAATCATGAAGCAATCGGCTAGAGGT
GAAATGAAGAGGACGTCAGAAAGAGGAGCTGGAAATCTGCCCTT
mm4_intergenic_Gm807|Zfp366_chr13_99159150 ACAGCCTTCAGAGACACTGCTCTGCCAGTGCACTGCTGTCCTGGGGGAGGAGCCAGTTGCTGTTTGTCATCTTGTGTAAC
CTTCAAGATTTGCATCTTCCTAGAGTTGAACCATTGCTCACTCCCAAAGGC
mm4_intergenic_Zc3h13|Siah3_chr14_75433050 TTAGGACAGTCACTGGCAGCTGCCAGCCAATTGCCTGGAGTGCTTAGACCATCACAAAATTGGATGGATTCCGACTCTAG
CATCAATCAGCTCCTTCTCCGGGAGGAAGGAAAATGCCCGTGGTG
mm4_intergenic_Csrnp1|Xirp1_chr9_119984668 TCTCTCCCGTTCTGACTGCANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNGTGGAAGCTCGACCCTCC
mm4_exon_Olfr380_chr11_73454164 GGACAGGAACAGGGCAAAGAACAGTTGCTGGTGCTCTGCAGGGATCGGCAGGCCCAGGAGGAGGAACTGGTTGAAGAGAG
TTTGGTTGTTCTTTATCATTCTTCCTCTCCA
mm4_intron_Vsig1_chrX_140918271 TGCCCATGCTGAGGAAGAACGGGCCTCCTGGAGGAGGAGGAGGAGGATGAAAAACACTTGCCAAAACACAGTTCCTTGTC
TTGTTTTCCCATCCTGTTTCAGTTTACCTACCCTTGCCATATCCCCATCACAGCCC
mm4_intergenic_Gm11367|Mboat1_chr13_30136442 GTCTCCCGGTCTCCTGCCCCGGGCCGGGCTCCCGGGGATCCGCGCCCATCTCTGCCCGGCTCCTCCTCCGGGGCGTCCCC
GGCTGCTGGCCAATGGGCGGCGGGCGGTGCGGAGCGGGAAGCAGAACGT
mm3_intron_Tnni3k_chr3_154811496 TGATGCGGCCTTAGTGTGTTTCTGAGTCCTAGGGTGACAGCTCAAGGAGTGAGCAAATCAGAATGCCCTGCATTCACAAT
CAAGCGGCTCCTCCTCCCAG
GCACCTCTGCTGTGATCTCACGCCTGTCA
mm4_intergenic_Glt1d1|Tmem132d_chr5_127756868 AACAAATGTGGTGCCATGGCAAGCTGCAGAGACACAAACCTGCCAGGAGGGGGAGCCAGTTGCTGCCATGTGCAGATGGG
TCCAGA
mm4_exon_Fam126a_chr5_23964406 GTAGCATCCCCACTGCTCTCACGTGAGGCTGTTTTCTCACATGCCTGTGGAGCAGCATGGCTTCATGGACTCGCTCCTCC
CCCTGG
GCTCTACAGCCCAGTCCTCTCACA
mm3_intergenic_Insig1|En2_chr5_28138257 TGCATGACTTTTCCCCACCTCTGTAGAATGCAGCTCCTGGTTCTACAACGATAGGCTCCTCCTCCTAGGGACCTCCTTAT
TCATGCCTCACTGTGGTGGATACAACCATGGCC
mm4_intergenic_RP24-86O15.2|Gm10698_chr9_33426252 GCATCGTACTGGGTTGTGGACTCAGAGAAAAGGTTCTGAACAATGTAGGTTTTTACCAGAGTAAAATGTCAATTAAAGAC
CTATAAATTCACTGCTGCTTCCACCCGTTCCTCCTCCAGGTCCCCACTTCAGCAATTGGA
mm4_intergenic_6530411M01Rik|Gm8492_chr17_9173834 CCTCAGACTGTTCCGAAGCACGGAGTCGGGGAAATAGATTAAAACCTGGAGGCTGTGAAGCCCCTGGAGGAAGACAGGGT
CGATG
CTCCACAACTGGTCTGAGAAACGGCT
mm3_exon_Incenp_chr19_9885741 TACCCATCTCTGGCCTGTGACCTTACCTCTTGCTCCTCTTCTGCCTCCTCAGGTTCGGCCACCTCGACAGGCTCCTGCTC
CAGG
CTGAAAGCAAAGCAGGAGCACTGTGAGAAC
mm4_intron_Prepl_chr17_85089247 ACTGATTCAGTGAACCGCCCAACTGACTGGCTCCTCCTCCTAGAGGAGCAATTGGATTTCACCA
mm4_intergenic_Rap1a|Adora3_chr3_105806971 AATCCTACCCAGGCCATTTACCACCCACCTGCTCCTCCCCCTGGAGATTGTTTCATGCCTTTGAAGTAAAAGTCAGCAGC
AGATTGGATTTTTAAAAACTGCTTCTTCACCGCAGACGCCGTATCTACCGAC
mm4_intergenic_Npm1|Gm12115_chr11_33163227 GCCTTTCATCCTCACGCCTAGGTACCTCGCCCCCCTCCTCCTCCAGGTGCTTGGCGGGCTCTCACTGCGCACGCGCGTTC
GTCTCTGGCCCCGTCCCTCCACTACTGTCGCCCAAG
mm4_exon_Espnl_chr1_91327564 TTCACAGACATCGGCCTGACAGCACGGGATAATGAAGGAGCCACTGCCCTGCACTTTGCAGCTCGGGGGGGCCACACACC
TATTCTCGACCGGCTCCTGCTCATGGGTGCCCCCATCATGAGAGACTCTTGGGGAGGG
mm4_intron_Lphn3_chr5_81062282 CAGCTTGTGTCCTGTTGCAGGAAGCAAACAACTTCTGGATAGAAAGGAAAATAAATAAGCAAACATCGTCTGGCTCCTTC
TCTTGG
AGGATTAATCTTCCATTACACTTCTTCCCACAGTTCTCAGGGGTTC
mm4_intergenic_Gm13375|Gm13377_chr2_21028402 TCCTGCTGGAGGAGCTGAAAGAAATGTGAAAAGGACGATATAGTTACAGAATATATTAAGCTTCTGGCTAGGATGGTATC
AAATGACCAGGAGGAGTATCCTGTAGATGCTGTTTATATAGGGATGTCACATGACCCCAG
mm4_intergenic_Vps13c|Gm22382_chr9_68126396 AGCCAAGAGTGAATCATGCTGATAAGCTGTATAGGAGATGATTATCCAGGAGGATGAGCAGGTAGATTGGGATGTTTATA
ATGCCATATGACAAAAGGTTAGCTCACAGCAGAAGGATCTTCTGGGAAGCAAGAAGGGC
mm4_exon_Rpusd3_chr6_113415625 TGCAGGGTCCGAGAGAAGTAAGGGGGCAGGGGTGCCAGAAGTTCACTAGGGGGGTCCCTGGATCCGGTGCCTGGGAGGAG
GAGGCGGTGAAGG
TGCAGGTGCAAAGGCATCTGGGCCACTTGGGAAGGACTTAGGCGG
mm3_intron_A830082K12Rik_chr13_78250814 TGACCCATGATGGTGTCTCTTTATAAGAACCCAGTCTCTGGGCCACCAGTTCTGGAGGAGGAGCCGTTTGATAGAGAGTA
TGACGTTTAGGACAAACATATTTATGAGGCCATTGGACACCTTT
mm4_exon_Spata22/Olfr20_chr11_73353777 CCAGCATGGCAGAAGGGAACCAAACTGTCATCTTCCAGTTCCTCCTCCTGGGCCTGCCCATCCCCACAGAGCATCAGCAG
CTGTACTATGCCCTGTTCCTGCTCATGTACCTCACCACCGTCC
mm4_exon_Fam20a_chr11_109722142 GCGTCTCTTCCAAGGATGCTGTCTCTCCGCAGCTGCTGAGACAGATCCAGCGGCTCCTCCTGCAGGCGGCAGGGACTCCT
GTGCCCCGGGCTGGCAGGAGCGGAGGGAACGCAGAGTTGACA
mm4_intron_Ifltd1_chr6_145613409 GTCCAAGGAGAGACCGGTTGGAGTAGGCTGTCCCAGGAGAGGGAGCAGGTCAATGCACTGTGTTGTGAATTTTAAAGAAA
ATTGTATGAATATGGATGAGGTGGGTGGCATTCT
mm4_exon_9130401M01Rik/Zhx1_chr15_58076377 GAGAGACTCCGCTCCCTGAACCCACAAGGGCCGCGAGGAGGAGCCAGCCGAGGCGGGGACGAGCCGGGCACCCCACTCGA
GGACTCC
mm4_exon_Mrm1_chr11_84819152 CTGCAGCCCAGCCTTACCGGCCTGGAGCAGGAGCCGGGCCACGCGCCGACGGGCAGCCCGCAAGGCCAGGAGGCATGGGG
ACAAGCCAAATAGACGTTCCAGCCGCT
mm4_exon_Olfr1_chr11_73395974 GGACGGTGGTGAGGTACATGACCAGGAACAGGGCATAGAACAGTTGCTGGTGCTCTGGTGGAATGGGTAGGCCCAGGAGG
AGGAACTGGGAGATG
ACAGTTTTGTTCCTTTCTGTCATAGTGTCT
mm4_intron_Slc45a1_chr4_150636752 TGCTCCTAGAAACAGCAGGCTGAACACCCTGGCGACTTGGACACTCCTCCCCAGGAGGAGCAGCCGCTCACTGCAGAACG
CTCTCCAGGAGAGGCTCAGCCAGGCGGGTGCATCTGTGGCCTGGGCAGAGTGTCTCACTG
mm4_exon_Ube2r2_chr4_41136125 CAGATGACCAGCTCGCAGAAAGCCCTGATGCTTGAGCTGAAATCCCTGCAGGAGGAACCGGTGGAGGGCTTCCGGATCAC
CCTGGTGGATGAGTCCGACCTCTACAACTGGGAGGTGGC
mm4_intergenic_Phlpp1|Bcl2/D630008O14Rik_chr1_106400614 CATTTGAGCTGGGCTTGCTGTAGTCCATCCACCTCCTGCTCCTCCAGGCAGGCTCCATCCTACAGTCCCCAGTACCCTCA
GA
mm4_intron_Pias1_chr9_62886113 CCAGATTGAAGCCCAAGGGTGTCTGTACGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNGTAAGTGTCTGGTGGCATCACA
mm4_exon_Olfr23_chr11_73940270 TGCCTGGAAAGAACCAAACTGTCATCTCCCGGTTCATCCTCCTGGGCCTGCCCATACCACCAGAGCACCAGCACCTGTTC
TATGCC
mm4_exon_Frrs1_chr3_116899140 GGCCGTCTTTGCACCATTTTCTCACCTGCCGGCTCCTCCTGCAGGTGCATCGGATGCTCATGGTAGCTACCTCCCTGCTC
ACCTGCGTTGCTTTTGTTTTGCCC
mm4_intron_Aqp3_chr4_41096442 CGGGCACTATGTCCAGTCAGACCCTGGCCTGAGCTCTTTGCCTGAGGCCCAGACTTGGCTCCGTGCCAAGGGGCCTCCGG
AGGAGGAGGCTGTCAAAG
CCAGTTCCACCACCTTGGGGCTAATC
mm4_intron_Oxct1_chr15_4135230 TTCGGCCTTTACAGAGAGCGGACGCTTCACCAGCACGAGGAGCCTGTCGAAGAGACAGGGCCTCAGAACCCTGCAGCTTC
TTCCCAGGGAACTGGCTGGGTCCATCCATTGTCTTAGTTGGCTGCTCTTCCCTTGAG

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.