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Contents:

PCR primers for off-targets of CTCCCGGGAAGTCGGAGGGC 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
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52313998_F Primer3: not found at this Tm N.d. 1.00
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52313998_R Primer3: not found at this Tm N.d. 1.00
mm4_intergenic_Hcrtr1|Tinagl1_chr4_130156886_F Primer3: not found at this Tm N.d. 0.55
mm4_intergenic_Hcrtr1|Tinagl1_chr4_130156886_R Primer3: not found at this Tm N.d. 0.55
mm4_exon_Atg7_chr6_114684439_F TCGTCGGCAGCGTCACTTCTACTACTGGTTTTGCTGC 58.9 0.48
mm4_exon_Atg7_chr6_114684439_R GTCTCGTGGGCTCGGGATCCAAGCTCACAGGTCCC 60.1 0.48
mm4_intron_Gna14_chr19_16537829_F TCGTCGGCAGCGTCAGAAGCTTGCCGCATTACCT 60.0 0.43
mm4_intron_Gna14_chr19_16537829_R GTCTCGTGGGCTCGGGATGCGTGCTCACTGTAGGT 60.1 0.43
mm4_intron_Osbpl1a_chr18_12840833_F Primer3: not found at this Tm N.d. 0.41
mm4_intron_Osbpl1a_chr18_12840833_R Primer3: not found at this Tm N.d. 0.41
mm3_exon_Phyhip_chr14_70467338_F TCGTCGGCAGCGTCAAGATCCCAGCTGCAAGACC 60.0 0.40
mm3_exon_Phyhip_chr14_70467338_R GTCTCGTGGGCTCGGTAAGCCACACATCACGTCCC 60.0 0.40
mm4_intron_Exoc4_chr6_33583321_F TCGTCGGCAGCGTCATTTCCTCTGCTACCTGCCC 59.4 0.40
mm4_intron_Exoc4_chr6_33583321_R GTCTCGTGGGCTCGGCTAACGAGTGCACTGCCAGA 60.0 0.40
mm4_intron_Tiam1_chr16_89928988_F Primer3: not found at this Tm N.d. 0.38
mm4_intron_Tiam1_chr16_89928988_R Primer3: not found at this Tm N.d. 0.38
mm3_exon_1190002N15Rik_chr9_94538065_F TCGTCGGCAGCGTCATCGGCTGTCTCCACAACTC 59.7 0.38
mm3_exon_1190002N15Rik_chr9_94538065_R GTCTCGTGGGCTCGGATAACTTGGCGGGCAAGGAG 60.3 0.38
mm4_intergenic_Efnb3|Wrap53_chr11_69560449_F TCGTCGGCAGCGTCCCCTTTGTCTCTGGCAGTGT 59.8 0.38
mm4_intergenic_Efnb3|Wrap53_chr11_69560449_R GTCTCGTGGGCTCGGAGCCTTTCTGGATGCTAGCG 60.1 0.38
mm4_intergenic_Chsy1/Gm26827|Lrrk1_chr7_66196774_F TCGTCGGCAGCGTCGCTGGGTGGCTGATAAGGTT 60.0 0.36
mm4_intergenic_Chsy1/Gm26827|Lrrk1_chr7_66196774_R GTCTCGTGGGCTCGGGGCAAGCTCTAAGACGACGA 59.8 0.36
mm4_intergenic_Ces2a|4932416K20Rik_chr8_104794344_F Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Ces2a|4932416K20Rik_chr8_104794344_R Primer3: not found at this Tm N.d. 0.33
mm4_intron_Lgr6_chr1_135021550_F TCGTCGGCAGCGTCAGTCTCCAAGCTTCATGCCC 60.0 0.32
mm4_intron_Lgr6_chr1_135021550_R GTCTCGTGGGCTCGGGTGGGTACAAGGTAGTTCAGGG 60.0 0.32
mm3_intron_Camk2g_chr14_20784765_F Primer3: not found at this Tm N.d. 0.31
mm3_intron_Camk2g_chr14_20784765_R Primer3: not found at this Tm N.d. 0.31
mm4_intergenic_Gm3571|Gm26239_chr10_93784544_F Primer3: not found at this Tm N.d. 0.29
mm4_intergenic_Gm3571|Gm26239_chr10_93784544_R Primer3: not found at this Tm N.d. 0.29
mm4_intergenic_Sox13|Gm23939_chr1_133492247_F Primer3: not found at this Tm N.d. 0.28
mm4_intergenic_Sox13|Gm23939_chr1_133492247_R Primer3: not found at this Tm N.d. 0.28
mm3_intergenic_Zdhhc25|Brd1_chr15_88629238_F TCGTCGGCAGCGTCAGGCATCCTGACATCCCTCT 60.0 0.25
mm3_intergenic_Zdhhc25|Brd1_chr15_88629238_R GTCTCGTGGGCTCGGGTCTGCATGCCTCCTGTTCT 60.0 0.25
mm3_intergenic_Tbx4|Gm11443_chr11_85916921_F Primer3: not found at this Tm N.d. 0.24
mm3_intergenic_Tbx4|Gm11443_chr11_85916921_R Primer3: not found at this Tm N.d. 0.24
mm4_intron_Micall1_chr15_79131609_F TCGTCGGCAGCGTCTTGTGTAGGAGCTGCTAGCC 59.4 0.23
mm4_intron_Micall1_chr15_79131609_R GTCTCGTGGGCTCGGCATTCTGTGACTTGGGGCCA 60.2 0.23
mm3_intergenic_Arfgef1|Cpa6_chr1_10238988_F Primer3: not found at this Tm N.d. 0.22
mm3_intergenic_Arfgef1|Cpa6_chr1_10238988_R Primer3: not found at this Tm N.d. 0.22
mm4_exon_Dnase2a/Mast1_chr8_84910761_F TCGTCGGCAGCGTCCCTACAGGTCCAGTTCTGGC 59.7 0.22
mm4_exon_Dnase2a/Mast1_chr8_84910761_R GTCTCGTGGGCTCGGGGTCCTCTGTGGCACTGAAA 59.8 0.22
mm4_intergenic_Gm22609|Lrrn2_chr1_132849260_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Gm22609|Lrrn2_chr1_132849260_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_A930031H19Rik|Gm25261_chr4_131182396_F Primer3: not found at this Tm N.d. 0.21
mm4_intergenic_A930031H19Rik|Gm25261_chr4_131182396_R Primer3: not found at this Tm N.d. 0.21
mm4_intergenic_Snora24|Ndst3_chr3_123508857_F TCGTCGGCAGCGTCTTTCTCGAACCGGGAGAACG 60.0 0.21
mm4_intergenic_Snora24|Ndst3_chr3_123508857_R GTCTCGTGGGCTCGGGCGGGAGTGAGATATGCCTT 59.6 0.21
mm4_intron_Rusc2_chr4_43397025_F TCGTCGGCAGCGTCTGCAACCCCTCTTTCTGTCC 59.8 0.21
mm4_intron_Rusc2_chr4_43397025_R GTCTCGTGGGCTCGGGTAACCCACTCTGTGCTCCC 60.0 0.21
mm4_intergenic_Tmie|Tmie/Als2cl_chr9_110878599_F TCGTCGGCAGCGTCATGAAGGAAGCTGGGATGCC 60.1 0.21
mm4_intergenic_Tmie|Tmie/Als2cl_chr9_110878599_R GTCTCGTGGGCTCGGTACTCTCCTGTGTACCCCGC 60.6 0.21
mm4_intron_Rpia_chr6_70778052_F Primer3: not found at this Tm N.d. 0.21
mm4_intron_Rpia_chr6_70778052_R Primer3: not found at this Tm N.d. 0.21
mm3_exon_D630003M21Rik_chr2_158220471_F TCGTCGGCAGCGTCACTTGCTCACACAGACGCTT 60.1 0.20
mm3_exon_D630003M21Rik_chr2_158220471_R GTCTCGTGGGCTCGGCCACTGTCCTGGGTCAAGTG 60.2 0.20
mm3_exon_D10Bwg1379e_chr10_18625208_F TCGTCGGCAGCGTCCAGCACTGTTCCCACTCAGA 59.6 0.19
mm3_exon_D10Bwg1379e_chr10_18625208_R GTCTCGTGGGCTCGGCCAGGAGCTTGTTCGGGAAT 60.0 0.19
mm3_intergenic_Gm26124|Gm16391_chr17_76147354_F Primer3: not found at this Tm N.d. 0.17
mm3_intergenic_Gm26124|Gm16391_chr17_76147354_R Primer3: not found at this Tm N.d. 0.17
mm4_intergenic_Trib2|mmu-mir-6387_chr12_15797047_F TCGTCGGCAGCGTCTTAACCAGGCCTGTGATGGG 59.6 0.17
mm4_intergenic_Trib2|mmu-mir-6387_chr12_15797047_R GTCTCGTGGGCTCGGACAGGGACTACACTACAGGT 57.0 0.17
mm4_intron_Gm11690_chr11_114233130_F Primer3: not found at this Tm N.d. 0.17
mm4_intron_Gm11690_chr11_114233130_R Primer3: not found at this Tm N.d. 0.17
mm4_intergenic_Wnt4|Cdc42_chr4_137311069_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Wnt4|Cdc42_chr4_137311069_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm23905|Zfp36l2_chr17_83979161_F TCGTCGGCAGCGTCGGTAGCCAACTGTGAGCAGA 59.6 0.15
mm4_intergenic_Gm23905|Zfp36l2_chr17_83979161_R GTCTCGTGGGCTCGGATCCCATTCCCCAGGGTGAT 60.3 0.15
mm4_exon_Tsc22d3_chrX_140542590_F TCGTCGGCAGCGTCTATTCGGCCTTCTCCTTGCC 59.8 0.15
mm4_exon_Tsc22d3_chrX_140542590_R GTCTCGTGGGCTCGGGCTGCTGCGTGGTGAAAA 59.2 0.15
mm4_intergenic_Rnf4|Fam193a_chr5_34369843_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Rnf4|Fam193a_chr5_34369843_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm23536|Gm4842_chr18_69241428_F TCGTCGGCAGCGTCTTCGCTGATGACATGCAGGT 60.0 0.14
mm4_intergenic_Gm23536|Gm4842_chr18_69241428_R GTCTCGTGGGCTCGGTGGCAAACACCCAGAACAGT 60.0 0.14
mm4_intergenic_Dusp8|Gm25370_chr7_142095882_F TCGTCGGCAGCGTCTGGTTTGCCTCAGGTCACTC 59.8 0.14
mm4_intergenic_Dusp8|Gm25370_chr7_142095882_R GTCTCGTGGGCTCGGCGTTTCCAAGCGGCTTTTCA 59.9 0.14
mm3_intergenic_Upf1|Comp_chr8_70358885_F Primer3: not found at this Tm N.d. 0.13
mm3_intergenic_Upf1|Comp_chr8_70358885_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Dok6|Gm22694_chr18_89624863_F Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Dok6|Gm22694_chr18_89624863_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_Mras|1600029I14Rik_chr9_99451715_F TCGTCGGCAGCGTCGAGCCTGGTGACCCGTTG 60.3 0.12
mm4_intergenic_Mras|1600029I14Rik_chr9_99451715_R GTCTCGTGGGCTCGGCAATCTCTGGGCAAAGCAGC 59.8 0.12
mm4_intron_Tulp4_chr17_6186030_F TCGTCGGCAGCGTCAGAGCCTTGCATCAGGGAAC 60.0 0.12
mm4_intron_Tulp4_chr17_6186030_R GTCTCGTGGGCTCGGTTACTCTACAGGGGAGGCCC 60.0 0.12
mm4_exon_AU022252/Gm12898_chr4_119224819_F TCGTCGGCAGCGTCGATCCCAGCAAAGCCCATCT 60.1 0.12
mm4_exon_AU022252/Gm12898_chr4_119224819_R GTCTCGTGGGCTCGGCTGTCCTGGTGTTCATGGCT 59.9 0.12
mm4_intergenic_Gm14397|Cdh22_chr2_165164463_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Gm14397|Cdh22_chr2_165164463_R Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Pfdn4/Cyp24a1|Gm16796_chr2_170497572_F TCGTCGGCAGCGTCGGCAAGGTTCCTGAGGACAG 60.3 0.11
mm4_intergenic_Pfdn4/Cyp24a1|Gm16796_chr2_170497572_R GTCTCGTGGGCTCGGAAAGGAAGTCCAGCTCAGGC 59.9 0.11
mm4_exon_Rps6kc1_chr1_190796730_F TCGTCGGCAGCGTCAAAGTACAGGCAGGTGGGTG 59.8 0.10
mm4_exon_Rps6kc1_chr1_190796730_R GTCTCGTGGGCTCGGGGAAAGTCCAGAGGGTGCTG 60.3 0.10
mm3_intron_Nomo1_chr7_46083343_F TCGTCGGCAGCGTCATATTGCCCTGCCTCTGACG 59.8 0.10
mm3_intron_Nomo1_chr7_46083343_R GTCTCGTGGGCTCGGACAACTTCCATGACACCGGG 60.2 0.10
mm4_intergenic_Gm24979|Cdc5l_chr17_45222119_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm24979|Cdc5l_chr17_45222119_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm10398|Gm26660_chr14_25170054_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm10398|Gm26660_chr14_25170054_R Primer3: not found at this Tm N.d. 0.09
mm4_intron_Tnni3k_chr3_154936774_F TCGTCGGCAGCGTCTGGATAGACATTTCTCTTCTCTGTT 57.5 0.09
mm4_intron_Tnni3k_chr3_154936774_R GTCTCGTGGGCTCGGACATGGGTGTGCACTATGCT 59.6 0.09
mm4_intergenic_Tfap2c|Gm22773_chr2_172728342_F TCGTCGGCAGCGTCTTATTGGCATCCTCGCTGGG 60.1 0.09
mm4_intergenic_Tfap2c|Gm22773_chr2_172728342_R GTCTCGTGGGCTCGGTGTGTTTAACTCCGCTCCCC 59.9 0.09
mm4_intergenic_Fmod|Btg2_chr1_134057167_F Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Fmod|Btg2_chr1_134057167_R Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Unc5b|mmu-mir-6408_chr10_60862400_F TCGTCGGCAGCGTCAACAACCCAGCCTAGGATGC 60.0 0.08
mm4_intergenic_Unc5b|mmu-mir-6408_chr10_60862400_R GTCTCGTGGGCTCGGGGTAGCAACTTCCTCCTGGG 59.7 0.08
mm4_intergenic_Gm8358|n-R5s63_chr12_82545558_F Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Gm8358|n-R5s63_chr12_82545558_R Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Ehd1|Cdc42bpg_chr19_6306396_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Ehd1|Cdc42bpg_chr19_6306396_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Brinp3|Gm15583_chr1_146702041_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Brinp3|Gm15583_chr1_146702041_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Slitrk1|Gm25901_chr14_109362907_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Slitrk1|Gm25901_chr14_109362907_R Primer3: not found at this Tm N.d. 0.07
mm4_exon_Mslnl_chr17_25745074_F Primer3: not found at this Tm N.d. 0.06
mm4_exon_Mslnl_chr17_25745074_R Primer3: not found at this Tm N.d. 0.06
mm4_exon_Tspyl3_chr2_153223013_F TCGTCGGCAGCGTCGAGCAAGGCCTTTCGTGAGA 60.3 0.06
mm4_exon_Tspyl3_chr2_153223013_R GTCTCGTGGGCTCGGCTCCTCCTCTGCCCCTCTAC 60.4 0.06
mm4_intergenic_Zfp622|March11_chr15_26135075_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Zfp622|March11_chr15_26135075_R Primer3: not found at this Tm N.d. 0.05
mm3_intron_Inadl_chr4_98699655_F Primer3: not found at this Tm N.d. 0.05
mm3_intron_Inadl_chr4_98699655_R Primer3: not found at this Tm N.d. 0.05
mm4_exon_Plekhh1_chr12_79074635_F Primer3: not found at this Tm N.d. 0.05
mm4_exon_Plekhh1_chr12_79074635_R Primer3: not found at this Tm N.d. 0.05
mm4_intron_1700025C18Rik_chr2_165079175_F TCGTCGGCAGCGTCGTATGATCACGGCCACCAGT 59.8 0.05
mm4_intron_1700025C18Rik_chr2_165079175_R GTCTCGTGGGCTCGGCCAAGAACCACACTCCACCA 59.8 0.05
mm4_intron_Rap1gap_chr4_137667735_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Rap1gap_chr4_137667735_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Kcng2|Ctdp1_chr18_80393991_F TCGTCGGCAGCGTCGTCTCCTTGTTCCCCAGGTG 59.9 0.04
mm4_intergenic_Kcng2|Ctdp1_chr18_80393991_R GTCTCGTGGGCTCGGCCTGTCCCTGGAGAGACACT 60.2 0.04
mm4_intergenic_Ndufc1|Naa15_chr3_51415887_F TCGTCGGCAGCGTCGAGCCAACTCTGAAATCCGC 59.2 0.04
mm4_intergenic_Ndufc1|Naa15_chr3_51415887_R GTCTCGTGGGCTCGGCTCTTCCCCACCCTCGATG 59.1 0.04
mm4_intron_Fez1_chr9_36869897_F Primer3: not found at this Tm N.d. 0.04
mm4_intron_Fez1_chr9_36869897_R Primer3: not found at this Tm N.d. 0.04
mm4_intron_Uck2_chr1_167273734_F TCGTCGGCAGCGTCAAGCCCAAGTACGAACAGCA 59.8 0.04
mm4_intron_Uck2_chr1_167273734_R GTCTCGTGGGCTCGGTGCTGAGCTTGTACACCCAC 60.2 0.04
mm4_intergenic_Olfr1387|Olfr1386_chr11_49469390_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Olfr1387|Olfr1386_chr11_49469390_R Primer3: not found at this Tm N.d. 0.04
mm4_exon_Brap_chr5_121660898_F Primer3: not found at this Tm N.d. 0.04
mm4_exon_Brap_chr5_121660898_R Primer3: not found at this Tm N.d. 0.04
mm4_intron_Jarid2_chr13_44851729_F TCGTCGGCAGCGTCACAACTGAGTGCTCCTGTGG 59.8 0.04
mm4_intron_Jarid2_chr13_44851729_R GTCTCGTGGGCTCGGGGGTGGAAGGTCAAGAATAAGGA 59.7 0.04
mm4_intergenic_Adora1/Mybph|Adora1_chr1_134228417_F TCGTCGGCAGCGTCTTAAGGTCCTGCCCCTCCAG 60.9 0.03
mm4_intergenic_Adora1/Mybph|Adora1_chr1_134228417_R GTCTCGTGGGCTCGGCCTTCCTCCCCTTTCGCATT 60.0 0.03
mm4_intron_Gas2_chr7_51869398_F TCGTCGGCAGCGTCCACCATGGTCACAGCAGGTA 59.6 0.03
mm4_intron_Gas2_chr7_51869398_R GTCTCGTGGGCTCGGTGTTCACAGCAGACCAGACA 59.1 0.03
mm3_intergenic_Gm23979|Gm22845_chr12_9653321_F Primer3: not found at this Tm N.d. 0.03
mm3_intergenic_Gm23979|Gm22845_chr12_9653321_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Limch1_chr5_66764363_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Limch1_chr5_66764363_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Kcnk12_chr17_87797099_F TCGTCGGCAGCGTCGCTCACCTATGGTCGACACC 60.1 0.03
mm4_exon_Kcnk12_chr17_87797099_R GTCTCGTGGGCTCGGCTTCCTCCGGCACTACGAG 59.5 0.03
mm4_intron_Dnah5_chr15_28356459_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Dnah5_chr15_28356459_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Bcs1l_chr1_74591828_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Bcs1l_chr1_74591828_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Gripap1_chrX_7804276_F TCGTCGGCAGCGTCTCACTCCTCTTCCTGCTTTTCC 59.9 0.02
mm4_intron_Gripap1_chrX_7804276_R GTCTCGTGGGCTCGGGTGGGCAAAAGGAACTGCAG 59.9 0.02
mm4_intergenic_Gabrg2|Gabra1_chr11_42044973_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gabrg2|Gabra1_chr11_42044973_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Agtr1a_chr13_30367445_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Agtr1a_chr13_30367445_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm8935|Gm2464_chr3_13142761_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm8935|Gm2464_chr3_13142761_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Tpd52_chr3_8954483_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Tpd52_chr3_8954483_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Bcl9l|Cxcr5_chr9_44511059_F TCGTCGGCAGCGTCGCCATGCACCAGATCTGTCT 60.1 0.02
mm4_intergenic_Bcl9l|Cxcr5_chr9_44511059_R GTCTCGTGGGCTCGGGCCAAAGGGGACAAAACACC 59.8 0.02
mm4_intron_Stpg2_chr3_139630771_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Stpg2_chr3_139630771_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Tpk1|Gm22939_chr6_43755370_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Tpk1|Gm22939_chr6_43755370_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Mc2r|mmu-mir-6356_chr18_68573427_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Mc2r|mmu-mir-6356_chr18_68573427_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm23859|Eif3s6-ps2_chr18_90265032_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm23859|Eif3s6-ps2_chr18_90265032_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Ankrd26|Cacna1c_chr6_118568266_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Ankrd26|Cacna1c_chr6_118568266_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm6395|Gm527_chr12_64896330_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm6395|Gm527_chr12_64896330_R Primer3: not found at this Tm N.d. 0.01
mm4_intron_Grhl2_chr15_37352500_F TCGTCGGCAGCGTCCAACTCAGCCCCAGTCACAG 60.6 0.01
mm4_intron_Grhl2_chr15_37352500_R GTCTCGTGGGCTCGGCCAGGTCGTGGTTCTCAAAC 58.7 0.01
mm4_intron_Clec10a_chr11_70165032_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Clec10a_chr11_70165032_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Tox3|Chd9_chr8_90713466_F TCGTCGGCAGCGTCATGACCCAAGGACCAGTTCC 59.3 0.01
mm4_intergenic_Tox3|Chd9_chr8_90713466_R GTCTCGTGGGCTCGGGCCATATGGGTCTGTGCCAT 60.1 0.01
mm4_intergenic_Gm22673|Gm23059_chr12_60619985_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm22673|Gm23059_chr12_60619985_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm16157|Fam169b_chr7_68275887_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Gm16157|Fam169b_chr7_68275887_R Primer3: not found at this Tm N.d. 0.01
mm3_intron_Mepe_chr5_104334102_F TCGTCGGCAGCGTCACGGCACTGTACTTCATGAGG 60.0 0.00
mm3_intron_Mepe_chr5_104334102_R GTCTCGTGGGCTCGGAGCCTGTCATAGTAACTGCGT 59.1 0.00
mm4_intron_Dmbx1_chr4_115929341_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Dmbx1_chr4_115929341_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Tst|Mpst_chr15_78406029_F TCGTCGGCAGCGTCTGGGAGAGGTTGGCACATTC 59.9 0.00
mm4_intergenic_Tst|Mpst_chr15_78406029_R GTCTCGTGGGCTCGGTCCTAGGCACCAGAAAGGGA 59.8 0.00
mm4_intron_Fmn1_chr2_113362011_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Fmn1_chr2_113362011_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Tex29|A230072I06Rik_chr8_12172185_F TCGTCGGCAGCGTCAGCTGGAGGAGGTTTTGACG 59.9 0.00
mm4_intergenic_Tex29|A230072I06Rik_chr8_12172185_R GTCTCGTGGGCTCGGCTGGGGAAGAAAGAGCATCCA 59.7 0.00
mm4_intergenic_Gnai1|Gm17037_chr5_18361959_F TCGTCGGCAGCGTCGCCCCTTCCCTTTGAAAAGC 59.6 0.00
mm4_intergenic_Gnai1|Gm17037_chr5_18361959_R GTCTCGTGGGCTCGGGGGCTCCAGTGCTCAGTTC 60.3 0.00
mm4_intergenic_Gm8036|Il1rapl2_chrX_137436330_F TCGTCGGCAGCGTCCTTTCCCCAGCTCAACACCT 59.8 0.00
mm4_intergenic_Gm8036|Il1rapl2_chrX_137436330_R GTCTCGTGGGCTCGGTGCTTGAGGAGGCACTTGAG 59.9 0.00
mm4_intergenic_Gm25011|Gm23680_chr10_12107142_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm25011|Gm23680_chr10_12107142_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm23795|Gm5342_chr7_69176187_F TCGTCGGCAGCGTCCCAGACCTTTGCACCTGTGA 60.1 0.00
mm4_intergenic_Gm23795|Gm5342_chr7_69176187_R GTCTCGTGGGCTCGGGGGCAAACTCTTTCCCTGGA 59.8 0.00
mm4_intergenic_Btla|Cd200_chr16_45287238_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Btla|Cd200_chr16_45287238_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Ankrd45|Prdx6_chr1_161232931_F TCGTCGGCAGCGTCGCTTAGTTCTTCTGCTCCTTGC 59.5 0.00
mm4_intergenic_Ankrd45|Prdx6_chr1_161232931_R GTCTCGTGGGCTCGGAGAGAAGGGCACGGCTAGTA 60.0 0.00
mm3_intergenic_Camta1|4930589P08Rik_chr4_151317934_F Primer3: not found at this Tm N.d. 0.00
mm3_intergenic_Camta1|4930589P08Rik_chr4_151317934_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_exon_Atg7_chr6_114684439 ACTTCTACTACTGGTTTTGCTGCCCCGCCCTCTGTCTTCCTGAGAGCATCCCTCTAATCCGGGGACCTGTGAGCTTGGAT
C
mm4_intron_Gna14_chr19_16537829 AGAAGCTTGCCGCATTACCTGGAAGCCCGTTCCAGAAGCTCCCCAGCCCTCTGGCTTCCAGGGACTGTGCTCGACAGCTT
TATGTAACTCACCTACAGTGAGCACGCATC
mm3_exon_Phyhip_chr14_70467338 AAGATCCCAGCTGCAAGACCTGCAACATCAGCGTGGGCCGCTAGGGACTCCTGGGGAGCCGGAGGGCAGGAGGGTGGGAC
GTGATGTGTGGCTTA
mm4_intron_Exoc4_chr6_33583321 ATTTCCTCTGCTACCTGCCCACTCTGTCTAACATCACCAGCACTCTGTCTTCCTGGGAGGTTGCATATGGCTGGTACACT
TCCTTTATGGAGACAGCACTTTGGTAACTTCTGTTTATCTGGCAGTGCACTCGTTAG
mm3_exon_1190002N15Rik_chr9_94538065 ATCGGCTGTCTCCACAACTCCCGGTGCCCTCCTCCGAGCTCCCCGCCCTCCGACTCCGCGGGCGCCGCCTCCTTGCCCGC
CAAGTTAT
mm4_intergenic_Efnb3|Wrap53_chr11_69560449 CCCTTTGTCTCTGGCAGTGTCCGTCTCTGGCTGAGTTTCTACTTGCTTTCCTTCCTGCCCTCCGATTGCCCGAGCGTTTG
GAATCTTCCTCGCTAGCATCCAGAAAGGCT
mm4_intergenic_Chsy1/Gm26827|Lrrk1_chr7_66196774 GCTGGGTGGCTGATAAGGTTATCTGGGGACTGTTTCTGGCAGTTCAAGTGCCTGTTGTACAACTTGGGGCTGGGGAAGTC
GGAGGGCTGG
GCAGGGCGGGGACTTCGTCGTCTTAGAGCTTGCC
mm4_intron_Lgr6_chr1_135021550 AGTCTCCAAGCTTCATGCCCCAGTCCCTCTGCCTTGTTAACTGTTTCCTTCTACGCATTATTTGATGTCTCTGCTAGCAG
GGACCCTGCCCTCCTACTTCCTGGGCTACCTACCCTGAACTACCTTGTACCCAC
mm3_intergenic_Zdhhc25|Brd1_chr15_88629238 AGGCATCCTGACATCCCTCTGCAGACCAGGCGGCCTCCCACCCTCCCTCCGCCTTCCTGGGAGCTGCTACTATTGGGGGT
TGGTTTTCAATCGCAAGGTCCATCCATGCTGGAGAACAGGAGGCATGCAGAC
mm4_intron_Micall1_chr15_79131609 TTGTGTAGGAGCTGCTAGCCCTGTCTCTGCCCCACGCCTGTCCTCAGACTGGTTCTTACCGTCTGTCTCTGTGACTGAAG
TATTACCTCTTCATTCACCCGGGAACTTGGAAGGCTGGAATGGCCCCAAGTCACAGAATG
mm4_exon_Dnase2a/Mast1_chr8_84910761 CCTACAGGTCCAGTTCTGGCAAAATTCTCCAGGCATCCTGCCCTCCAACTGCTCTGGAGCCTATCAGGTTCTGGATGTGA
CACAGACAGGATTCCCTGGCCCATCTAGACTAACTTTCAGTGCCACAGAGGACC
mm4_intergenic_Snora24|Ndst3_chr3_123508857 TTTCTCGAACCGGGAGAACGGGAAGCCTGGGCAGTGCGAGCAGACCCGCCCGGCCCTCGGCTTTTCCGGGAGGAAGGCAT
ATCTCACTCCCGC
mm4_intron_Rusc2_chr4_43397025 TGCAACCCCTCTTTCTGTCCTGACCCATAGCATCCAGGGAAGTCAGAGGGAGGGATGTCCCCGAGGACTTGAAAACTGGG
AGTGGAGGGAAAGGCCCTGTGGGAGCACAGAGTGGGTTAC
mm4_intergenic_Tmie|Tmie/Als2cl_chr9_110878599 ATGAAGGAAGCTGGGATGCCCCTTTCTTCCTCCCAGGCAGGCAGAGGGCAGGGGTTAGAGAAGTAGGTATCTGGGGGATG
GTGCATCCTCGGGGGTAGGGGAGGTGGGAGCGGGGTACACAGGAGAGTA
mm3_exon_D630003M21Rik_chr2_158220471 ACTTGCTCACACAGACGCTTGGCAGGGATGAGAAAATCCAAGAGGAAGCTCAGTCCATCTCCCCGGAAGTCGGAGTCCAG
G
AGCCGGAACACTTGACCCAGGACAGTGG
mm3_exon_D10Bwg1379e_chr10_18625208 CAGCACTGTTCCCACTCAGACTACCTCCCCGGAAGTCCGAGGTCCGGCCACGGCTACATTCCCGAACAAGCTCCTGG
mm4_intergenic_Trib2|mmu-mir-6387_chr12_15797047 TTAACCAGGCCTGTGATGGGGAAGGAGAGACAACAGGAATGAAACCCTCCCTCCACCTTCTCGGGAGGAACTCCACAGAA
TTTATGCTATGCCAAATCTTTAAACCTGTAGTGTAGTCCCTGT
mm4_intergenic_Gm23905|Zfp36l2_chr17_83979161 GGTAGCCAACTGTGAGCAGACAGCAAGACTGGGAAGTGTACTTTCTGCCTCTGAAACAACCTGCCCCCATCAGGAGGCTA
AACTCCAGGGAGGTCAGAGGTCGGGGAATTTTTTTTTGGATCACCCTGGGGAATGGGAT
mm4_exon_Tsc22d3_chrX_140542590 TATTCGGCCTTCTCCTTGCCCCAGCCTTGCCTAGCTCGCCGGCCCTCTGCCTTCCCGGCTGCAGCTGCTGTTTTTCACCA
CGCAGCAGC
mm4_intergenic_Gm23536|Gm4842_chr18_69241428 TTCGCTGATGACATGCAGGTGTGCACGGGTGTGGTACTCAGTCTTGACGCTTTCGTGTTCTCCTGGGTGGTTGGAGGGCC
GG
CTCCATTAGTTTGTGGCCTTGTGAGAATATATAAAACTGTTCTGGGTGTTTGCCA
mm4_intergenic_Dusp8|Gm25370_chr7_142095882 TGGTTTGCCTCAGGTCACTCATCCCGGGAAATCAGGGGGCGGGGCTCCAGACCCACCGGTGTGCTCCCCAGGGCGGCGCT
GCCCCCCTACCTATGACATCCAGTTCTTGACAGCTTCCAATGAAAAGCCGCTTGGAAACG
mm4_intergenic_Mras|1600029I14Rik_chr9_99451715 GAGCCTGGTGACCCGTTGGCTCCCGGGAGGAAGGAGGACAGGAGGGAGAGCAAGAGTTAGTCTATGTCTTTGTCATTTTG
CATCAGTACACTGATAGAACCTCTCTGGCCTCGCTGCTTTGCCCAGAGATTG
mm4_intron_Tulp4_chr17_6186030 AGAGCCTTGCATCAGGGAACACACTCCAGAGAAGACTGAGGGCAGGAAGGGTGGGCTGCAGTCACAGGCTTTGTATGTGG
TGGTGAAAGTGGGCCTCCCCTGTAGAGTAA
mm4_exon_AU022252/Gm12898_chr4_119224819 GATCCCAGCAAAGCCCATCTTCCAGCCCTGCAGCTTCCAGGGAGGGCGGTTTCTGCACGTCCCTTGTTAGATCTCAGTTC
TTGTGCTAGGCATGGATGTTTCTAGCTCGTAAGTGAAGCCATGAACACCAGGACAG
mm4_intergenic_Pfdn4/Cyp24a1|Gm16796_chr2_170497572 GGCAAGGTTCCTGAGGACAGTGCGGAAGACCGCCCGCGCTTGCCAAGAGAGTGGAGAGCGAGCCTGCCTCCCAACATCCC
GGGAG
ACAAGTTTTCGTAGCCTGAGCTGGACTTCCTTT
mm4_exon_Rps6kc1_chr1_190796730 AAAGTACAGGCAGGTGGGTGATGGTTTTTGAACAAACAGCTGATAAGGATCCCGGGAGGGAGGAGGGCAGGGGCAGGCCA
CACTGGATAAACAGAGGGCAGAGCTCCCCTCAGCACCCTCTGGACTTTCC
mm3_intron_Nomo1_chr7_46083343 ATATTGCCCTGCCTCTGACGCTGCTGCTCCTGTTGGCGGGCTACAACCATGACAAGGTAGGGAAGCCAAGGCCTCATGGG
AAGTCGGGGGGCAGG
AGATAGGCTATGGGAGGGTGCCCGGTGTCATGGAAGTTGT
mm4_intron_Tnni3k_chr3_154936774 TGGATAGACATTTCTCTTCTCTGTTATGATGTGAAATCTCCTGGGAAGTTGGAGAGTGGGGCATGTATAAATTTCATTTT
GCACATGGAAATGTACCATATGCAATCATCTGTAAGCATAGTGCACACCCATGT
mm4_intergenic_Tfap2c|Gm22773_chr2_172728342 TTATTGGCATCCTCGCTGGGCCCTGTGTGAGAACGCGCCGGCCTGGCCCTGCAGAGGGGCCAACGTCTATTACATTCCAA
TAAGTGTCGCCCGGGAGGGAGGAGGGCGGGGCGGGGAGCGGAGTTAAACACA
mm4_intergenic_Unc5b|mmu-mir-6408_chr10_60862400 AACAACCCAGCCTAGGATGCCCACTGAAAGCTCAAACCCAAGCCTCTGAACCTGAAAACTGCCCCCGTGCCAGCCATTCT
GCTTCCCGGGAG
AGAATGGGGCTCCCAGGAGGAAGTTGCTACC
mm4_exon_Tspyl3_chr2_153223013 GAGCAAGGCCTTTCGTGAGATGACAACCTGGGGGGAACTCGGAGGGCAGGGCAGTTTAGCCTTTTATCTATAGGAACCCA
CAAGGGGTAGAGGGGCAGAGGAGGAG
mm4_intron_1700025C18Rik_chr2_165079175 GTATGATCACGGCCACCAGTTAATTAGTGATCAATGGCCTACAGGGAAGTGAGAGGGCGGGTGGCTGTGGTGGAGTGTGG
TTCTTGG
mm4_intergenic_Kcng2|Ctdp1_chr18_80393991 GTCTCCTTGTTCCCCAGGTGCAAATCTGAACTGACTTTTCTGGAACAAGCCATCTTCTCCAGGGAAGTGGGAAGTCAGGA
AGCAGCATCCTGGGAAGTGACCAGGATGCCTTAGTGTCTCTCCAGGGACAGG
mm4_intergenic_Ndufc1|Naa15_chr3_51415887 GAGCCAACTCTGAAATCCGCGGCTCCGCAGGCCGGCCCGGCCTCCGCGTTCCCGGCAGGCTCCGGTGCATCGAGGGTGGG
GAAGAG
mm4_intron_Uck2_chr1_167273734 AAGCCCAAGTACGAACAGCACTATCCATATCTCCCAGCAGTCAATGAGACAAACAGGCTGAAATGCTCAACCAGTGTCTG
AGGGCTCCAGGGCAGTTGGGGGGCGGGGGGGTGGGTGTACAAGCTCAGCA
mm4_intron_Jarid2_chr13_44851729 ACAACTGAGTGCTCCTGTGGGGTTTTTGTTTATATGTTAATTAATTGAAAGAGGTTGTTCAGGCTGTCTTTGACCCTGCC
CTCAGCCTTCCCAGCAG
TGTAATTAGTTATGTGTCCTTATTCTTGACCTTCCACCC
mm4_intergenic_Adora1/Mybph|Adora1_chr1_134228417 TTAAGGTCCTGCCCCTCCAGCTCGGAAGAGCAAATAGGATGATAAATACCAGATGCCTCCTGGGAAGTCGCAGAGTAGGT
GAAATGCGAAAGGGGAGGAAGG
mm4_intron_Gas2_chr7_51869398 CACCATGGTCACAGCAGGTAGACAAAGGTTTCTTAAGCAAAGCTCTCCCAACCCTCCCACTTCCCGGTAAAGTTCTCTCA
CTTTGGGTTTGGAGTGCTACTCATTTCAGTATTCTGTCTGGTCTGCTGTGAACA
mm4_exon_Kcnk12_chr17_87797099 GCTCACCTATGGTCGACACCACGGTGCCCACGAAGTAGAAGGCTCCAGGGAAGTCCCAGCGCGGGCGCAGCGCGTCGGCG
CGGACCCCGGCGGCCAGCGCGGCCTCGTAGTGCCGGAGGAAG
mm4_intron_Gripap1_chrX_7804276 TCACTCCTCTTCCTGCTTTTCCATTTTAAACGTTGTCTTTCTTCCTCCCTTTATACTTCTCTCTCTGTCTCCCCGGAACT
GGGAGGTCCGG
GGTTCCTGGGCCTTAGGGCCCCCTGCAGTTCCTTTTGCCCAC
mm4_intergenic_Bcl9l|Cxcr5_chr9_44511059 GCCATGCACCAGATCTGTCTATGCCCACCAGACACAGCCACTGCCTCTCCTCTGACTTCCCAGGAGAGCCCCTCACCCGC
CCCTGGTCAGACTCAAGCCATACAGGAATGGGTGTTTTGTCCCCTTTGGC
mm4_intron_Grhl2_chr15_37352500 CAACTCAGCCCCAGTCACAGCCTTTGCTTCTGGCCCCACCCCCTGTGGCCTCCCCTCAGATTTCCTGGGAGTGGATGAGC
AATCACAAAAACAGCAAGTTCTTTGCTTGAACACTGATGTTTGAGAACCACGACCTGG
mm4_intergenic_Tox3|Chd9_chr8_90713466 ATGACCCAAGGACCAGTTCCAACTCTCACGGGAGGGCGGAGGGGGGGGGGGGGCTGTATCTTTTAGAAGTTACAGAAAGT
CAGTCTGGGAGAAAATGGCACAGACCCATATGGC
mm3_intron_Mepe_chr5_104334102 ACGGCACTGTACTTCATGAGGGGGTTTCTGAGCATATTCTAGACCATATGCTAGAACAGTGTAGACTCCCGGGGAGTCGG
GGGGGGGG
GAACGCAGTTACTATGACAGGCT
mm4_intergenic_Tst|Mpst_chr15_78406029 TGGGAGAGGTTGGCACATTCCCTAGGGAAAGCCTGCTCCACCCCTCCCACTTCCCCGGCGCCCTCCCTTTCTGGTGCCTA
GGA
mm4_intergenic_Tex29|A230072I06Rik_chr8_12172185 AGCTGGAGGAGGTTTTGACGGCTCCCTGGAAGACGGCGGGAAGGTGGATGCTCTTTCTTCCCCAG
mm4_intergenic_Gnai1|Gm17037_chr5_18361959 GCCCCTTCCCTTTGAAAAGCTCGTCAAGGACATGAGGAAATAATCCTCACTTCTGAGAGGATGTTGGAAGAACTCTCCAG
GGAAGCTGGTGGGCAGG
GAACTGAGCACTGGAGCCC
mm4_intergenic_Gm8036|Il1rapl2_chrX_137436330 CTTTCCCCAGCTCAACACCTGCCTTGCCACTCCTGGGAAGTAGGTTGGCTGGTTGCAATTAAACTCAAGTGCCTCCTCAA
GCA
mm4_intergenic_Gm23795|Gm5342_chr7_69176187 CCAGACCTTTGCACCTGTGAAAGGCACATTTGTTCCCAGGGATGCCCTCCCTGGCCCTTCTCCCAGGAAGTGGGCAGGCA
GG
GTGGGTGTCAGTTTTCTGTGTCTAGTTGCTCCAGGGAAAGAGTTTGCCC
mm4_intergenic_Ankrd45|Prdx6_chr1_161232931 GCTTAGTTCTTCTGCTCCTTGCATCCCGGGAGGCCGGTGGGCAGGCTGGTCTTAAGTAGTGATCCTCAGTATAGGTGTGA
AGGCAATCCAAGGCTTACTAGCCGTGCCCTTCTCT

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.