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Contents:

PCR primers for off-targets of CCAAGTAAGCTCCTAATCCC AGG

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_52313785_F Primer3: not found at this Tm N.d. 1.00
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52313785_R Primer3: not found at this Tm N.d. 1.00
mm4_intergenic_mmu-mir-6393|Zdhhc25_chr15_87925947_F Primer3: not found at this Tm N.d. 0.68
mm4_intergenic_mmu-mir-6393|Zdhhc25_chr15_87925947_R Primer3: not found at this Tm N.d. 0.68
mm3_intergenic_Veph1|Shox2_chr3_66423976_F Primer3: not found at this Tm N.d. 0.43
mm3_intergenic_Veph1|Shox2_chr3_66423976_R Primer3: not found at this Tm N.d. 0.43
mm4_intron_9330182L06Rik_chr5_9391273_F Primer3: not found at this Tm N.d. 0.32
mm4_intron_9330182L06Rik_chr5_9391273_R Primer3: not found at this Tm N.d. 0.32
mm4_intergenic_RP23-448C3.1/Trpm3|Trpm3_chr19_22467463_F Primer3: not found at this Tm N.d. 0.31
mm4_intergenic_RP23-448C3.1/Trpm3|Trpm3_chr19_22467463_R Primer3: not found at this Tm N.d. 0.31
mm4_intergenic_Eda2r|Gm9062_chrX_97785674_F Primer3: not found at this Tm N.d. 0.27
mm4_intergenic_Eda2r|Gm9062_chrX_97785674_R Primer3: not found at this Tm N.d. 0.27
mm4_intergenic_Kcnk12|Rpl36-ps4_chr17_87879197_F TCGTCGGCAGCGTCCCTTGGACCCTTCTGCCTTT 59.8 0.25
mm4_intergenic_Kcnk12|Rpl36-ps4_chr17_87879197_R GTCTCGTGGGCTCGGACACTATACAGAGGGAGGGTCC 60.0 0.25
mm4_intergenic_Kbtbd8|AY512915_chr6_95221215_F Primer3: not found at this Tm N.d. 0.24
mm4_intergenic_Kbtbd8|AY512915_chr6_95221215_R Primer3: not found at this Tm N.d. 0.24
mm4_intron_Dgkb_chr12_37886106_F Primer3: not found at this Tm N.d. 0.23
mm4_intron_Dgkb_chr12_37886106_R Primer3: not found at this Tm N.d. 0.23
mm4_intergenic_4631405K08Rik|Plxna2_chr1_193722503_F Primer3: not found at this Tm N.d. 0.20
mm4_intergenic_4631405K08Rik|Plxna2_chr1_193722503_R Primer3: not found at this Tm N.d. 0.20
mm4_intergenic_C330008G21Rik|Ptprt_chr2_162626694_F Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_C330008G21Rik|Ptprt_chr2_162626694_R Primer3: not found at this Tm N.d. 0.19
mm4_intron_Magi2_chr5_20472457_F Primer3: not found at this Tm N.d. 0.18
mm4_intron_Magi2_chr5_20472457_R Primer3: not found at this Tm N.d. 0.18
mm4_intron_Sept8_chr11_53525599_F TCGTCGGCAGCGTCTGCTCTCTCCCATACGACCT 59.7 0.17
mm4_intron_Sept8_chr11_53525599_R GTCTCGTGGGCTCGGCTTAGGCTCTGGTCCCAGGA 60.3 0.17
mm4_intergenic_Nek7|Lhx9_chr1_138771300_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Nek7|Lhx9_chr1_138771300_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm22708|Zbtb46_chr2_181435692_F TCGTCGGCAGCGTCCAGGACCAAGGGAGCAGAAA 59.5 0.15
mm4_intergenic_Gm22708|Zbtb46_chr2_181435692_R GTCTCGTGGGCTCGGGAGATTAAGCAGGGCAGCCT 59.8 0.15
mm3_intron_Gpr64_chrX_160409468_F Primer3: not found at this Tm N.d. 0.15
mm3_intron_Gpr64_chrX_160409468_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Ighv14-4|Ighv3-3_chr12_114193849_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Ighv14-4|Ighv3-3_chr12_114193849_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm12700|Cachd1_chr4_100749553_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm12700|Cachd1_chr4_100749553_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm25423|Gm26054_chr7_116716283_F TCGTCGGCAGCGTCGCCACAACCCTCAGTCTTCA 59.8 0.14
mm4_intergenic_Gm25423|Gm26054_chr7_116716283_R GTCTCGTGGGCTCGGCCCTGCCCTAAAGCATCACT 59.7 0.14
mm4_intron_Plcl1_chr1_55736496_F TCGTCGGCAGCGTCTGCAGCTCTACATGGAAGCA 59.3 0.12
mm4_intron_Plcl1_chr1_55736496_R GTCTCGTGGGCTCGGCCTGTTCTGGGCTTGGGTAG 60.0 0.12
mm4_intron_Clock_chr5_76298162_F Primer3: not found at this Tm N.d. 0.12
mm4_intron_Clock_chr5_76298162_R Primer3: not found at this Tm N.d. 0.12
mm4_intron_Col6a1_chr10_76712680_F TCGTCGGCAGCGTCTGCTTCAGCCTGTTGATGGT 59.8 0.11
mm4_intron_Col6a1_chr10_76712680_R GTCTCGTGGGCTCGGGTTTCCCTTCCCAGTGAGCA 59.8 0.11
mm4_intron_D330045A20Rik_chrX_139537235_F Primer3: not found at this Tm N.d. 0.11
mm4_intron_D330045A20Rik_chrX_139537235_R Primer3: not found at this Tm N.d. 0.11
mm4_exon_Zfy2_chrY_2133460_F Primer3: not found at this Tm N.d. 0.10
mm4_exon_Zfy2_chrY_2133460_R Primer3: not found at this Tm N.d. 0.10
mm4_exon_Zfy1_chrY_759925_F TCGTCGGCAGCGTCTCATCCATGGCCTTCAGTTCT 59.0 0.10
mm4_exon_Zfy1_chrY_759925_R GTCTCGTGGGCTCGGTGAGTGCCTCAGAGATGCAA 59.0 0.10
mm4_intergenic_Gm5803|Gm19840_chr15_22827561_F TCGTCGGCAGCGTCTCTTCTTCTCTACCACAGCCT 57.8 0.10
mm4_intergenic_Gm5803|Gm19840_chr15_22827561_R GTCTCGTGGGCTCGGTCTGTGATGCCCAAAAGCCA 60.1 0.10
mm3_intergenic_Neurog1|Cxcl14_chr13_56259422_F Primer3: not found at this Tm N.d. 0.09
mm3_intergenic_Neurog1|Cxcl14_chr13_56259422_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Esrrg_chr1_187710621_F Primer3: not found at this Tm N.d. 0.09
mm4_intron_Esrrg_chr1_187710621_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm25659|Gm25644_chr19_51197077_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm25659|Gm25644_chr19_51197077_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Adam30|Reg4_chr3_98219627_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Adam30|Reg4_chr3_98219627_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Sfxn5_chr6_85310087_F Primer3: not found at this Tm N.d. 0.09
mm4_intron_Sfxn5_chr6_85310087_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm22536|Gm6632_chr5_58462721_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm22536|Gm6632_chr5_58462721_R Primer3: not found at this Tm N.d. 0.09
mm3_intron_Rad17_chr13_100623874_F Primer3: not found at this Tm N.d. 0.08
mm3_intron_Rad17_chr13_100623874_R Primer3: not found at this Tm N.d. 0.08
mm4_intron_Ank1_chr8_23110162_F TCGTCGGCAGCGTCTCACCTCCTTTCTCCCCACT 59.8 0.08
mm4_intron_Ank1_chr8_23110162_R GTCTCGTGGGCTCGGCTGACCGTCATACCACCACC 60.1 0.08
mm4_intron_Gm14033_chr2_55428819_F TCGTCGGCAGCGTCCAGGTCCTCTCAGAAGCAGC 60.1 0.08
mm4_intron_Gm14033_chr2_55428819_R GTCTCGTGGGCTCGGCCTGGTCAGAGAAAGCCTGG 60.0 0.08
mm4_intron_Slc36a2_chr11_55178293_F Primer3: not found at this Tm N.d. 0.07
mm4_intron_Slc36a2_chr11_55178293_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Gm21865|Gm21882_chrY_40352225_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Gm21865|Gm21882_chrY_40352225_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Gm21943|Gm21724_chrY_58021491_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21943|Gm21724_chrY_58021491_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21918|Gm21832_chrY_61037318_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21918|Gm21832_chrY_61037318_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21916|Gm21756_chrY_33180599_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21916|Gm21756_chrY_33180599_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21914|Gm21852_chrY_40518214_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21914|Gm21852_chrY_40518214_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21861|Gm21758_chrY_58888303_F TCGTCGGCAGCGTCCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21861|Gm21758_chrY_58888303_R GTCTCGTGGGCTCGGAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21842|Gm21744_chrY_32322058_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21842|Gm21744_chrY_32322058_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21822|Gm21763_chrY_18911680_F TCGTCGGCAGCGTCGCACAGGCAATCCAAGGTTC 59.7 0.06
mm4_intergenic_Gm21822|Gm21763_chrY_18911680_R GTCTCGTGGGCTCGGTCTGGGGTTATACATGGTGGT 58.1 0.06
mm4_intergenic_Gm21805|Gm21241_chrY_42873097_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21805|Gm21241_chrY_42873097_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21803|Gm20837_chrY_90336261_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21803|Gm20837_chrY_90336261_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21780|Gm21737_chrY_31307400_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21780|Gm21737_chrY_31307400_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21779|Gm21247_chrY_45891024_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21779|Gm21247_chrY_45891024_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21767|Gm20822_chrY_14199410_F TCGTCGGCAGCGTCGCACAGGCAATCCAAGGTTC 59.7 0.06
mm4_intergenic_Gm21767|Gm20822_chrY_14199410_R GTCTCGTGGGCTCGGTCTGGGGTTATACATGGTGGT 58.1 0.06
mm4_intergenic_Gm21758|Gm21843_chrY_59695244_F TCGTCGGCAGCGTCCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21758|Gm21843_chrY_59695244_R GTCTCGTGGGCTCGGAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21751|Gm21271_chrY_48246923_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21751|Gm21271_chrY_48246923_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21745|Gm21184_chrY_34226362_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21745|Gm21184_chrY_34226362_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21739|Gm21856_chrY_44410439_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21739|Gm21856_chrY_44410439_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21739|Gm21856_chrY_44321891_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21739|Gm21856_chrY_44321891_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21724|Gm21405_chrY_58508785_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21724|Gm21405_chrY_58508785_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21679|Gm21529_chrY_30812193_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21679|Gm21529_chrY_30812193_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21679|Gm21529_chrY_29749771_F TCGTCGGCAGCGTCCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21679|Gm21529_chrY_29749771_R GTCTCGTGGGCTCGGAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21247|Gm21256_chrY_47385998_F TCGTCGGCAGCGTCAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21247|Gm21256_chrY_47385998_R GTCTCGTGGGCTCGGCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21247|Gm21256_chrY_46832972_F TCGTCGGCAGCGTCCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21247|Gm21256_chrY_46832972_R GTCTCGTGGGCTCGGAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21170|Gm21751_chrY_47752955_F TCGTCGGCAGCGTCCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm21170|Gm21751_chrY_47752955_R GTCTCGTGGGCTCGGAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm21094|Gm21588_chrY_32685432_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm21094|Gm21588_chrY_32685432_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm20901|Gm21865_chrY_39389564_F TCGTCGGCAGCGTCGGCACAGGCAATCCAAGGAT 60.6 0.06
mm4_intergenic_Gm20901|Gm21865_chrY_39389564_R GTCTCGTGGGCTCGGTCTGGGGTTATACATGGTGGT 58.1 0.06
mm4_intergenic_Gm20901|Gm21865_chrY_38960203_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm20901|Gm21865_chrY_38960203_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm20838|Gm21626_chrY_33608101_F TCGTCGGCAGCGTCCTTCTCTCATCGTCTGCGGT 59.5 0.06
mm4_intergenic_Gm20838|Gm21626_chrY_33608101_R GTCTCGTGGGCTCGGAAGGATCACCATGCTTCCCA 58.9 0.06
mm4_intergenic_Gm20810|Gm21853_chrY_29134561_F TCGTCGGCAGCGTCGGCACAGGCAATCCAAGGAT 60.6 0.06
mm4_intergenic_Gm20810|Gm21853_chrY_29134561_R GTCTCGTGGGCTCGGTCTGGGGTTATACATGGTGGT 58.1 0.06
mm4_intergenic_Gm20794|Gm21918_chrY_60543337_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm20794|Gm21918_chrY_60543337_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_1700040F15Rik|Gm21562_chrY_31708385_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_1700040F15Rik|Gm21562_chrY_31708385_R Primer3: not found at this Tm N.d. 0.06
mm4_intron_Ercc6_chr14_32536440_F TCGTCGGCAGCGTCTGAGACCTCCTGTGTGCCTA 59.8 0.06
mm4_intron_Ercc6_chr14_32536440_R GTCTCGTGGGCTCGGTCCTTTGCCTTAGAGGAAGCTG 60.0 0.06
mm4_intron_Itih5_chr2_10225553_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Itih5_chr2_10225553_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Shprh|Gm16577/Shprh_chr10_11210231_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Shprh|Gm16577/Shprh_chr10_11210231_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Rarb|Thrb_chr14_16797504_F TCGTCGGCAGCGTCTTCTGCTCTGGTCTGTCTGC 59.6 0.05
mm4_intergenic_Rarb|Thrb_chr14_16797504_R GTCTCGTGGGCTCGGGGGACTGGGAAATCACGACA 59.6 0.05
mm4_intergenic_Fau-ps1|Macrod2_chr2_141944759_F TCGTCGGCAGCGTCTGAGAGTTTTCTTATGGCAGCA 57.9 0.04
mm4_intergenic_Fau-ps1|Macrod2_chr2_141944759_R GTCTCGTGGGCTCGGTCTTGCATTCGCTGACCCTT 59.9 0.04
mm4_intergenic_Gm8730|Cdh5_chr8_103588461_F TCGTCGGCAGCGTCACACTTGTCTCCCAGGGACT 60.1 0.04
mm4_intergenic_Gm8730|Cdh5_chr8_103588461_R GTCTCGTGGGCTCGGGGAGCTAGGCTTTCTCTTGCA 60.0 0.04
mm4_intron_Fat3_chr9_16349244_F TCGTCGGCAGCGTCCCTGTCTGTGAGTGTTAAACCC 58.8 0.04
mm4_intron_Fat3_chr9_16349244_R GTCTCGTGGGCTCGGAGAGTCTCCTGCAGCTACCA 59.9 0.04
mm4_intergenic_Cadm1|Nxpe2_chr9_48290502_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Cadm1|Nxpe2_chr9_48290502_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Il10ra_chr9_45254414_F TCGTCGGCAGCGTCGTATAGGGAGGCAGGGACCA 60.1 0.03
mm4_exon_Il10ra_chr9_45254414_R GTCTCGTGGGCTCGGCTTCCACCTCATGCCAGTGT 59.9 0.03
mm4_intergenic_Gm23558|Galc_chr12_98009523_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm23558|Galc_chr12_98009523_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Ythdf3|Gm26485_chr3_16709680_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Ythdf3|Gm26485_chr3_16709680_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Nhsl1_chr10_18429079_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Nhsl1_chr10_18429079_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Cnot6|Gm12192_chr11_49690580_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Cnot6|Gm12192_chr11_49690580_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm6531|Olfr371_chr8_85174380_F TCGTCGGCAGCGTCGCACACTGGGACAAAGAGGA 59.8 0.03
mm4_intergenic_Gm6531|Olfr371_chr8_85174380_R GTCTCGTGGGCTCGGTGAGAAAATGATAATGAAGGCCACT 58.8 0.03
mm4_exon_Olfr1154_chr2_87903223_F Primer3: not found at this Tm N.d. 0.03
mm4_exon_Olfr1154_chr2_87903223_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Pde7b|Ahi1_chr10_20908803_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Pde7b|Ahi1_chr10_20908803_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm14544|4930525M21Rik/4930525M21RIK_chrX_37952860_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm14544|4930525M21Rik/4930525M21RIK_chrX_37952860_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Sp4|Gm25577_chr12_118468574_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Sp4|Gm25577_chr12_118468574_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Lonp2_chr8_86705363_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Lonp2_chr8_86705363_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_A730085K08Rik|9530059O14Rik_chr9_122499548_F TCGTCGGCAGCGTCCAGCAGTGATGATGGAAAGCC 59.5 0.01
mm4_intergenic_A730085K08Rik|9530059O14Rik_chr9_122499548_R GTCTCGTGGGCTCGGAGGCTATGTGAACAGCCTGG 59.7 0.01
mm4_intron_Vrk1_chr12_106034049_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Vrk1_chr12_106034049_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Opcml|Ntm_chr9_28944990_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Opcml|Ntm_chr9_28944990_R Primer3: not found at this Tm N.d. 0.01
mm4_intron_Dync1i1_chr6_5893610_F TCGTCGGCAGCGTCAGGGACTTCATGTTTGAGCCA 59.5 0.01
mm4_intron_Dync1i1_chr6_5893610_R GTCTCGTGGGCTCGGTCGCAGTCTGATTCTGAACGA 59.4 0.01
mm4_intergenic_Scarb1|Ubc_chr5_125364091_F TCGTCGGCAGCGTCGCCAGCGCATACTGCTCTAT 60.3 0.01
mm4_intergenic_Scarb1|Ubc_chr5_125364091_R GTCTCGTGGGCTCGGAACCATTGAACCATGCCCCA 60.1 0.01
mm4_intergenic_Cdh7|Cdh19_chr1_110166439_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Cdh7|Cdh19_chr1_110166439_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Rgs6|Gm22149_chr12_82694809_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Rgs6|Gm22149_chr12_82694809_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Wdpcp_chr11_21714072_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Wdpcp_chr11_21714072_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tyw1_chr5_130319584_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tyw1_chr5_130319584_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tpm1_chr9_67032157_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tpm1_chr9_67032157_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Slc9a2_chr1_40734902_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Slc9a2_chr1_40734902_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Pnpla7_chr2_25021415_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Pnpla7_chr2_25021415_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Glis3_chr19_28406025_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Glis3_chr19_28406025_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Cwf19l1_chr19_44127036_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Cwf19l1_chr19_44127036_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Col22a1_chr15_71950279_F TCGTCGGCAGCGTCACCCAGATCGAAGGAGCAAA 59.0 0.00
mm4_intron_Col22a1_chr15_71950279_R GTCTCGTGGGCTCGGCTGCCCAGGGAGAAGTCAAC 60.3 0.00
mm4_intergenic_Xrcc4|Gm4117_chr13_89856412_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Xrcc4|Gm4117_chr13_89856412_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Pvrl3|Gm6912_chr16_46589145_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Pvrl3|Gm6912_chr16_46589145_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Myl9|Gm14231_chr2_156783833_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Myl9|Gm14231_chr2_156783833_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Lpl|Slc18a1_chr8_69014012_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Lpl|Slc18a1_chr8_69014012_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Kcna6|Kcna6/Gm26673_chr6_126732832_F TCGTCGGCAGCGTCTTAAGGCAGCAGTTCCAGCA 59.8 0.00
mm4_intergenic_Kcna6|Kcna6/Gm26673_chr6_126732832_R GTCTCGTGGGCTCGGGCTTCTGTGATGCAGGCTTG 59.8 0.00
mm4_intergenic_Gm23061|Ostf1_chr19_18547124_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm23061|Ostf1_chr19_18547124_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm22307|Gm6085_chr1_4574280_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm22307|Gm6085_chr1_4574280_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm15941|4930447A16Rik_chr15_37438986_F TCGTCGGCAGCGTCTCCCTGTCATCCAAGGTTTCC 59.6 0.00
mm4_intergenic_Gm15941|4930447A16Rik_chr15_37438986_R GTCTCGTGGGCTCGGGCCAACAAGTTTGCATAGGGG 60.0 0.00
mm4_intergenic_Gm11999|Ikzf1_chr11_11658649_F TCGTCGGCAGCGTCAGTTATGTGCCAGACAGATGCA 60.0 0.00
mm4_intergenic_Gm11999|Ikzf1_chr11_11658649_R GTCTCGTGGGCTCGGACATACCACTGCACCCAAGG 59.9 0.00
mm4_intergenic_1700001G01Rik|Gm26235_chr18_17908665_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_1700001G01Rik|Gm26235_chr18_17908665_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Tgif2/5430405H02Rik_chr2_156853642_F TCGTCGGCAGCGTCTACACACTCCGCTGCCTTTC 60.3 0.00
mm4_exon_Tgif2/5430405H02Rik_chr2_156853642_R GTCTCGTGGGCTCGGAATGGTCCTGATGCTTGGCA 59.9 0.00
mm4_exon_Gm7148_chr17_40116063_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Gm7148_chr17_40116063_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Gm12605_chr4_89217808_F TCGTCGGCAGCGTCTGCCACCTTTGCTACACACT 59.8 0.00
mm4_exon_Gm12605_chr4_89217808_R GTCTCGTGGGCTCGGGCTTGGCAGTGATCCGTACT 60.1 0.00
mm3_intron_Tmcc1_chr6_116101377_F Primer3: not found at this Tm N.d. 0.00
mm3_intron_Tmcc1_chr6_116101377_R Primer3: not found at this Tm N.d. 0.00
mm3_intergenic_Trim29|Pvrl1_chr9_43427196_F TCGTCGGCAGCGTCTGCTGGGCTTGCTTTCTTTG 59.6 0.00
mm3_intergenic_Trim29|Pvrl1_chr9_43427196_R GTCTCGTGGGCTCGGAAACTGGCCCATACTCTGGC 60.0 0.00
mm3_intergenic_Gm10051|Gm15627_chr5_134067821_F Primer3: not found at this Tm N.d. 0.00
mm3_intergenic_Gm10051|Gm15627_chr5_134067821_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_Kcnk12|Rpl36-ps4_chr17_87879197 CCTTGGACCCTTCTGCCTTTGTAGCAGCATGGTGATCAGCTGCGTGCCTCACGGGTGATTCAGGCCCACCTGGGAATAGA
GACCATGAAAGCTCCTGACCCCTGGCAGGGACCCTCCCTCTGTATAGTGT
mm4_intron_Sept8_chr11_53525599 TGCTCTCTCCCATACGACCTTTTAGTTATTTCCCGTGACCTCTGACCTGGGTTTAGAGGCTTGCTTGGAATCCTGGGACC
AGAGCCTAAG
mm4_intergenic_Gm22708|Zbtb46_chr2_181435692 CAGGACCAAGGGAGCAGAAAGTTCTGTGACTCACAAATTCAAGCTGTGGCATTTGTGACAGTCATACGACACTTCTACCA
AGTAGCCACCTAATACCAGG
GAAGCAAAGGCTGCCCTGCTTAATCTC
mm4_intergenic_Gm25423|Gm26054_chr7_116716283 GCCACAACCCTCAGTCTTCATGTGTTTCCATCTGTACTCCCTGGGATTAGCCTCTTGCTTGGCTCTCTTGTGTACAGGGT
CTCCTGGCTTTGGGACTTTTCATTTACTCTGCTGTCTCTGAGTGATGCTTTAGGGCAGGG
mm4_intron_Plcl1_chr1_55736496 TGCAGCTCTACATGGAAGCAGAGGAATGGACCAAATGGCTTCCCAAGGATGTTTCTAATCCCTGGAGCCCAAGATGCACT
GGCTCTACCCAAGCCCAGAACAGG
mm4_intron_Col6a1_chr10_76712680 TGCTTCAGCCTGTTGATGGTTACTTGAAACATTAGGGAGAAAGGCCAGGGATCAGCAGCTTCCTAGGAGGCCTGACCCCA
GGGCACTGCTCACTGGGAAGGGAAAC
mm4_exon_Zfy1_chrY_759925 TCATCCATGGCCTTCAGTTCTTACTAAGTCAGCTCCTAAACCAGGGAACAACAAATACTGNNNNNNNNNNNNNNNNNNNN
NGTCATATTTGCATCTCTGAGGCACTCA
mm4_intergenic_Gm5803|Gm19840_chr15_22827561 TCTTCTTCTCTACCACAGCCTATTTCATTTAAGCTTCTAATCCCAGGTTCTTTGTCATTTCTTTCTTCAATATAAATTAA
TCTTGGCTTTTGGGCATCACAGA
mm4_intron_Ank1_chr8_23110162 TCACCTCCTTTCTCCCCACTCCCAAGTAAGCCCCAAATCAAGGGAACCCAGGATAGCGAGGATGGGCTTGATCTGTGGCC
CCCTCTGTCTAGAATTTGGTGGTGGTATGACGGTCAG
mm4_intron_Gm14033_chr2_55428819 CAGGTCCTCTCAGAAGCAGCTCTTCCGCTGGTGGATGCCAGTTATTAGCAGCTTAGTTGGGTTTAAGGTCCAGGCTTTCT
CTGACCAGG
mm4_intergenic_Gm21918|Gm21832_chrY_61037318 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21916|Gm21756_chrY_33180599 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21861|Gm21758_chrY_58888303 CTTCTCTCATCGTCTGCGGTTATACATGGTGATATTTATGAGACCTCTAAATGATCCCTAGGATTAGGAGCATAGATGGT
AGAGTTGGGAGTGGATATATAGACTGGAGAGATAAAATGGGAAGCATGGTGATCCTT
mm4_intergenic_Gm21842|Gm21744_chrY_32322058 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21822|Gm21763_chrY_18911680 GCACAGGCAATCCAAGGTTCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATG
CTCCTAATCCTAGG
GATCATTTAGAGGTCTCATAAATACCACCATGTATAACCCCAGA
mm4_intergenic_Gm21805|Gm21241_chrY_42873097 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21803|Gm20837_chrY_90336261 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21780|Gm21737_chrY_31307400 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21767|Gm20822_chrY_14199410 GCACAGGCAATCCAAGGTTCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATG
CTCCTAATCCTAGG
GATCATTTAGAGGTCTCATAAATACCACCATGTATAACCCCAGA
mm4_intergenic_Gm21758|Gm21843_chrY_59695244 CTTCTCTCATCGTCTGCGGTTATACATGGTGATATTTATGAGACCTCTAAATGATCCCTAGGATTAGGAGCATAGATGGT
AGAGTTGGGAGTGGATATATAGACTGGAGAGATAAAATGGGAAGCATGGTGATCCTT
mm4_intergenic_Gm21751|Gm21271_chrY_48246923 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21745|Gm21184_chrY_34226362 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21724|Gm21405_chrY_58508785 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21679|Gm21529_chrY_29749771 CTTCTCTCATCGTCTGCGGTTATACATGGTGATATTTATGAGACCTCTAAATGATCCCTAGGATTAGGAGCATAGATGGT
AGAGTTGGGAGTGGATATATAGACTGGAGAGATAAAATGGGAAGCATGGTGATCCTT
mm4_intergenic_Gm21247|Gm21256_chrY_47385998 AAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTATGCTCCTAATCCTAG
G
GATCATTTAGAGGTCTCATAAATATCACCATGTATAACCGCAGACGATGAGAGAAG
mm4_intergenic_Gm21247|Gm21256_chrY_46832972 CTTCTCTCATCGTCTGCGGTTATACATGGTGATATTTATGAGACCTCTAAATGATCCCTAGGATTAGGAGCATAGATGGT
AGAGTTGGGAGTGGATATATAGACTGGAGAGATAAAATGGGAAGCATGGTGATCCTT
mm4_intergenic_Gm21170|Gm21751_chrY_47752955 CTTCTCTCATCGTCTGCGGTTATACATGGTGATATTTATGAGACCTCTAAATGATCCCTAGGATTAGGAGCATAGATGGT
AGAGTTGGGAGTGGATATATAGACTGGAGAGATAAAATGGGAAGCATGGTGATCCTT
mm4_intergenic_Gm20901|Gm21865_chrY_39389564 GGCACAGGCAATCCAAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTAT
GCTCCTAATCCTAGG
GATCATTTAGAGGTCTCATAAATACCACCATGTATAACCCCAGA
mm4_intergenic_Gm20838|Gm21626_chrY_33608101 CTTCTCTCATCGTCTGCGGTTATACATGGTGATATTTATGAGACCTCTAAATGATCCCTAGGATTAGGAGCATAGATGGT
AGAGTTGGGAGTGGATATATAGACTGGAGAGATAAAATGGGAAGCATGGTGATCCTT
mm4_intergenic_Gm20810|Gm21853_chrY_29134561 GGCACAGGCAATCCAAGGATCACCATGCTTCCCATTTTATCTCTCCAGTCTATATATCCACTCCCAACTCTACCATCTAT
GCTCCTAATCCTAGG
GATCATTTAGAGGTCTCATAAATACCACCATGTATAACCCCAGA
mm4_intron_Ercc6_chr14_32536440 TGAGACCTCCTGTGTGCCTAGGACGTCCTAGGATTAGGATGGTACTTGGTAGACCAGATGCCTAAGTCTACTTGACCAGG
AACAGCTTCCTCTAAGGCAAAGGA
mm4_intergenic_Rarb|Thrb_chr14_16797504 TTCTGCTCTGGTCTGTCTGCCCCTACCCCTTCTCCATGCCCCTACTTCCTTTTTGCCCAAATAACCTCCTTATACCAGGT
GTATTGCATGTCGTGATTTCCCAGTCCC
mm4_intergenic_Fau-ps1|Macrod2_chr2_141944759 TGAGAGTTTTCTTATGGCAGCATAACTGGGAGCATAATTAACCATGAAGACACAGTGTATTTCCATTTTAGCTTCTAATC
CCAGG
AGATACCATGTTTTAGAGCTATGAGTAATTAAAGGGTCAGCGAATGCAAGA
mm4_intergenic_Gm8730|Cdh5_chr8_103588461 ACACTTGTCTCCCAGGGACTCTTGACCTTCAAGTTCTCTCCTTCAGAGCTGATCCAGGTTAGCTCCTAAACCTGGGAAAA
AAATCTAAAATCCAAAGACGTATTGTTAACCCTGAGATTTGCAAGAGAAAGCCTAGCTCC
mm4_intron_Fat3_chr9_16349244 CCTGTCTGTGAGTGTTAAACCCAATTTAAGGCAGAGATTAGAAGCCATCATATTACATTACCTATAGAAAAACCCAGGTG
CCCAAGAATGTTCCTAATCGCGGGTCCAGGAAGTGATGGTAGCTGCAGGAGACTCT
mm4_exon_Il10ra_chr9_45254414 GTATAGGGAGGCAGGGACCAGCCCTAGACCCCAGGGACTAGCAGCTCCCTTGGCACAAGGCCCTAGGCTCCCACAAGATC
TCTGAAACACTGGCATGAGGTGGAAG
mm4_intergenic_Gm6531|Olfr371_chr8_85174380 GCACACTGGGACAAAGAGGAAGCACACTTTCTCAGCAGAGAATCTCGATGTTCTGCCAAGTAAGCTGCTTACCCAAGGAA
GTGGCCTTCATTATCATTTTCTCA
mm4_intergenic_A730085K08Rik|9530059O14Rik_chr9_122499548 CAGCAGTGATGATGGAAAGCCAAAATTCCCAAGAAAACTCCTCAGCCCAGGGACTATGTCCAGCCCAGGCTGTTCACATA
GCCT
mm4_intron_Dync1i1_chr6_5893610 AGGGACTTCATGTTTGAGCCAATGGGAAAACATGAAGTTAACACTAGGGGAAATGACTTAGCCCCCAATAAAGCTGCTAA
TGCCAGG
ATAAAAGCAAACAAATCTGTTAATGATCTTCGTTCAGAATCAGACTGCGA
mm4_intergenic_Scarb1|Ubc_chr5_125364091 GCCAGCGCATACTGCTCTATATCCTGCCATTAGGAACTTACTTCGCTAGCACAGTGTCCTGGCAGTCTGCCCACTGCAGT
CTGGGTCAAATGTCCTTTCTTCAAGAGTGAGTGAGGACTGGGGCATGGTTCAATGGTT
mm4_intron_Col22a1_chr15_71950279 ACCCAGATCGAAGGAGCAAATGAGAGCTAAGAAATTATATACCTTTCCAAGTAAGCTGTGAGTCCCAGGCACCAACTAGG
CCTTTTGAGTTCTAATCAAGGGATCTTGGGTCTGGAAATGTTGACTTCTCCCTGGGCAG
mm4_intergenic_Kcna6|Kcna6/Gm26673_chr6_126732832 TTAAGGCAGCAGTTCCAGCAGACGGCTCAGGGAGCCTGGCATCTCCTGGGAATGGGATCTTTCTTGGCATTTACATGGAG
ATGCAAAGTGAGAGAAATAGCACTGCCAGCCAAGCCTGCATCACAGAAGC
mm4_intergenic_Gm15941|4930447A16Rik_chr15_37438986 TCCCTGTCATCCAAGGTTTCCTAACAAGCAAGCTTCTATTCCCTGGCAACCAAAGTTACACTGAATAATGGTTAAAAATG
ACAGATCCCCTATGCAAACTTGTTGGC
mm4_intergenic_Gm11999|Ikzf1_chr11_11658649 AGTTATGTGCCAGACAGATGCAGCCAAGTAAGCTATGAACCCCAGGATATGTTATCATTGATGGGATAATGGCTCTTAAA
GCCTTGGGTGCAGTGGTATGT
mm4_exon_Tgif2/5430405H02Rik_chr2_156853642 TACACACTCCGCTGCCTTTCGTCTCAGAAAACGCCAAGTAGGCACCTGCTCCCGGGTGGCTTGAGGTTCCTGGCTCACTG
CCCTTTCATGCAGAGGGTTATCCAAGTACGGATCACTGCCAAGCATCAGGACCATT
mm4_exon_Gm12605_chr4_89217808 TGCCACCTTTGCTACACACTCCTATGCCTCTCGTCTCAGAAAACGCCAAGTAGGCACCTGCTCCCGGGTGGCTTGAGGTT
CCTGGCTCACTGCCCTTTCATACAGAGGGTTATCCAAGTACGGATCACTGCCAAGC
mm3_intergenic_Trim29|Pvrl1_chr9_43427196 TGCTGGGCTTGCTTTCTTTGTGTGCTTGCTCAGTTACCAAGTTAGCTCGTATTCCCTGGCAAGCACTGGGCAGCCAGAGT
ATGGGCCAGTTT

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.