← return to the list of all guides

Contents:

PCR primers for off-targets of CGCCGGATGCGGGAGGAAGG CGG

In the table below, Illumina Nextera Handle sequences have been added and highlighted in bold. Primers for the on-target have been added for convenience. The table below is sorted by the CFD off-target score. Sites with very low CFD scores < 0.02 are unlikely to be cleaved, see our study Haeussler et al. 2016, Figure 2.

In the protocol by Matthew Canver, Harvard, two PCRs are run: one PCR to amplify the potential off-target, then a second PCR to extend the handles with Illumina barcodes. Please click here to download the protocol. Alternatively, you can have a look at Fu et al, 2014.

If a primer was not found, the reason is usually that the region around the off-target is too repetitive. To avoid unspecific primers, all repeats are masked for the primer design (not for off-target search). If you think that we should change the parameters here or should use different primer3 settings, please let us know.

Maximum amplicon length:     Primer Tm:

NamePrimer Sequence Tm CFD Score
mm0_exon_Pop5_chr5_115238018_F TCGTCGGCAGCGTCGAGCGTGGAGATTGACAGGG 60.4 1.00
mm0_exon_Pop5_chr5_115238018_R GTCTCGTGGGCTCGGTTACCTGTGCTTGAACCGCA 60.1 1.00
mm4_intergenic_Alg10b|Cpne8_chr15_90461789_F Primer3: not found at this Tm N.d. 0.65
mm4_intergenic_Alg10b|Cpne8_chr15_90461789_R Primer3: not found at this Tm N.d. 0.65
mm4_intergenic_Tnp2|Prm3_chr16_10790235_F Primer3: not found at this Tm N.d. 0.52
mm4_intergenic_Tnp2|Prm3_chr16_10790235_R Primer3: not found at this Tm N.d. 0.52
mm4_intergenic_Kntc1|Niacr1_chr5_123824989_F Primer3: not found at this Tm N.d. 0.51
mm4_intergenic_Kntc1|Niacr1_chr5_123824989_R Primer3: not found at this Tm N.d. 0.51
mm4_intergenic_Fgfr2|Gm5903_chr7_130279582_F TCGTCGGCAGCGTCTATGGGTCCCTCCTACAGGC 60.1 0.50
mm4_intergenic_Fgfr2|Gm5903_chr7_130279582_R GTCTCGTGGGCTCGGGCCCAGAAGAAACAGGAGCT 59.9 0.50
mm4_exon_Kpna1_chr16_36000258_F Primer3: not found at this Tm N.d. 0.50
mm4_exon_Kpna1_chr16_36000258_R Primer3: not found at this Tm N.d. 0.50
mm4_intron_Ptprn2_chr12_116486724_F TCGTCGGCAGCGTCGTGGATGGTAGGCAGTGAGG 59.8 0.49
mm4_intron_Ptprn2_chr12_116486724_R GTCTCGTGGGCTCGGCCTCCTCTCTGCAGTCCCTT 60.6 0.49
mm4_intergenic_Myh9|Txn2_chr15_77888701_F TCGTCGGCAGCGTCACAGAGCTCCAGGTAGAGGG 60.0 0.45
mm4_intergenic_Myh9|Txn2_chr15_77888701_R GTCTCGTGGGCTCGGCCGATCAGCCCTTTCTCTCC 59.8 0.45
mm4_intron_Grm7_chr6_111500187_F TCGTCGGCAGCGTCAGCCAAGATGCTGTTCCTCA 59.3 0.42
mm4_intron_Grm7_chr6_111500187_R GTCTCGTGGGCTCGGGGCATCTTCCAGTTTTCCATGT 59.1 0.42
mm4_intergenic_Gm10620|Cmc2_chr8_116863675_F TCGTCGGCAGCGTCGACATCCCCTGCAGATGGAG 59.8 0.38
mm4_intergenic_Gm10620|Cmc2_chr8_116863675_R GTCTCGTGGGCTCGGCGCGAGGGTCTAGTCTTCTC 59.3 0.38
mm4_intergenic_Begain|Gm22234_chr12_109147018_F Primer3: not found at this Tm N.d. 0.37
mm4_intergenic_Begain|Gm22234_chr12_109147018_R Primer3: not found at this Tm N.d. 0.37
mm4_exon_Cklf_chr8_104263916_F Primer3: not found at this Tm N.d. 0.36
mm4_exon_Cklf_chr8_104263916_R Primer3: not found at this Tm N.d. 0.36
mm3_intergenic_Gm3839|Ptprg_chr14_11552736_F TCGTCGGCAGCGTCTTTGCCCTCTGCCTAACCAG 59.9 0.34
mm3_intergenic_Gm3839|Ptprg_chr14_11552736_R GTCTCGTGGGCTCGGAAGGTCTGTCGCGGAGTTTT 59.8 0.34
mm4_intergenic_Gm23752|Gm24815_chr12_7420266_F Primer3: not found at this Tm N.d. 0.34
mm4_intergenic_Gm23752|Gm24815_chr12_7420266_R Primer3: not found at this Tm N.d. 0.34
mm4_intergenic_Mdfic|Tfec_chr6_16545293_F Primer3: not found at this Tm N.d. 0.32
mm4_intergenic_Mdfic|Tfec_chr6_16545293_R Primer3: not found at this Tm N.d. 0.32
mm4_exon_Angel2_chr1_190928833_F Primer3: not found at this Tm N.d. 0.32
mm4_exon_Angel2_chr1_190928833_R Primer3: not found at this Tm N.d. 0.32
mm4_intron_Tet2_chr3_133510561_F Primer3: not found at this Tm N.d. 0.31
mm4_intron_Tet2_chr3_133510561_R Primer3: not found at this Tm N.d. 0.31
mm4_intron_4930465A12Rik_chr4_128155355_F Primer3: not found at this Tm N.d. 0.31
mm4_intron_4930465A12Rik_chr4_128155355_R Primer3: not found at this Tm N.d. 0.31
mm4_intergenic_Diap2|Gm14984_chrX_130389026_F TCGTCGGCAGCGTCGCAACCCTACCTGTGCTCTC 60.3 0.30
mm4_intergenic_Diap2|Gm14984_chrX_130389026_R GTCTCGTGGGCTCGGGTTCACACAACTAAGCCATGGT 59.1 0.30
mm4_intron_Maea_chr5_33365609_F TCGTCGGCAGCGTCTGGCCTGCCAGAAGAAAACT 59.8 0.29
mm4_intron_Maea_chr5_33365609_R GTCTCGTGGGCTCGGATGGCCTAGGATCCACAGGT 60.0 0.29
mm4_intergenic_CT030684.1|Gm23276_chr14_97311670_F Primer3: not found at this Tm N.d. 0.28
mm4_intergenic_CT030684.1|Gm23276_chr14_97311670_R Primer3: not found at this Tm N.d. 0.28
mm4_intergenic_Cd52|Ubxn11_chr4_134095082_F Primer3: not found at this Tm N.d. 0.28
mm4_intergenic_Cd52|Ubxn11_chr4_134095082_R Primer3: not found at this Tm N.d. 0.28
mm4_exon_Brix1_chr15_10475665_F Primer3: not found at this Tm N.d. 0.22
mm4_exon_Brix1_chr15_10475665_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Lalba|Olfr279_chr15_98491078_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Lalba|Olfr279_chr15_98491078_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Nfia|Nfia/E130114P18Rik_chr4_97775148_F TCGTCGGCAGCGTCGCTGTAGTTGGAACTGGGCT 59.9 0.21
mm4_intergenic_Nfia|Nfia/E130114P18Rik_chr4_97775148_R GTCTCGTGGGCTCGGCAAGTTCAGCACCAAAGCCC 59.9 0.21
mm4_intergenic_F730016J06Rik|Olfm1_chr2_28189175_F TCGTCGGCAGCGTCACAAGGTTAGAGGAGGGGCA 60.1 0.21
mm4_intergenic_F730016J06Rik|Olfm1_chr2_28189175_R GTCTCGTGGGCTCGGAAGGCTAGCAGCTAGTGTGC 60.1 0.21
mm4_exon_Gsk3b_chr16_38089430_F Primer3: not found at this Tm N.d. 0.20
mm4_exon_Gsk3b_chr16_38089430_R Primer3: not found at this Tm N.d. 0.20
mm3_intergenic_Agpat9|Gm20484_chr5_101361707_F Primer3: not found at this Tm N.d. 0.19
mm3_intergenic_Agpat9|Gm20484_chr5_101361707_R Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Gm2366/Fat1|Gm2366_chr8_45020786_F TCGTCGGCAGCGTCAGGAGGAAAGAGAACACGTGT 58.9 0.19
mm4_intergenic_Gm2366/Fat1|Gm2366_chr8_45020786_R GTCTCGTGGGCTCGGTTCGTTCACCAGAATGCCCC 60.6 0.19
mm4_intergenic_Gm14601/Gm15482|Gm14644_chrX_54091434_F Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Gm14601/Gm15482|Gm14644_chrX_54091434_R Primer3: not found at this Tm N.d. 0.18
mm4_intron_Epb4.1l3_chr17_69242615_F Primer3: not found at this Tm N.d. 0.18
mm4_intron_Epb4.1l3_chr17_69242615_R Primer3: not found at this Tm N.d. 0.18
mm4_intergenic_Gm24686|Zfp804b_chr5_6743193_F Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Gm24686|Zfp804b_chr5_6743193_R Primer3: not found at this Tm N.d. 0.16
mm4_intergenic_Foxp1|Gm24936_chr6_99522897_F TCGTCGGCAGCGTCCTCTCAAAGCCCCTGTGACC 60.3 0.16
mm4_intergenic_Foxp1|Gm24936_chr6_99522897_R GTCTCGTGGGCTCGGCCTCTCTTGCGCGCCTTATT 60.8 0.16
mm4_intergenic_PVT1_2|Pvt1_chr15_62095843_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_PVT1_2|Pvt1_chr15_62095843_R Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm10076|Gm22406_chr14_105798288_F Primer3: not found at this Tm N.d. 0.15
mm4_intergenic_Gm10076|Gm22406_chr14_105798288_R Primer3: not found at this Tm N.d. 0.15
mm4_intron_Gm26791_chr14_118369252_F Primer3: not found at this Tm N.d. 0.14
mm4_intron_Gm26791_chr14_118369252_R Primer3: not found at this Tm N.d. 0.14
mm4_exon_Prss41/Prss32_chr17_23843721_F Primer3: not found at this Tm N.d. 0.14
mm4_exon_Prss41/Prss32_chr17_23843721_R Primer3: not found at this Tm N.d. 0.14
mm3_intron_Usp6nl_chr2_6323263_F Primer3: not found at this Tm N.d. 0.13
mm3_intron_Usp6nl_chr2_6323263_R Primer3: not found at this Tm N.d. 0.13
mm4_intron_Baiap2l2_chr15_79268641_F TCGTCGGCAGCGTCGTGGCTCTATTAAGGATGTTTCAAA 57.6 0.13
mm4_intron_Baiap2l2_chr15_79268641_R GTCTCGTGGGCTCGGCCCTGCAGAAGTGAGTTCCA 59.6 0.13
mm4_intron_Bri3bp_chr5_125443865_F Primer3: not found at this Tm N.d. 0.13
mm4_intron_Bri3bp_chr5_125443865_R Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_2310050C09Rik|Lce3a_chr3_92893518_F Primer3: not found at this Tm N.d. 0.13
mm4_intergenic_2310050C09Rik|Lce3a_chr3_92893518_R Primer3: not found at this Tm N.d. 0.13
mm4_intron_Acad12_chr5_121614594_F TCGTCGGCAGCGTCGACCCCAATCTCCTGCCTTC 60.1 0.13
mm4_intron_Acad12_chr5_121614594_R GTCTCGTGGGCTCGGACATCTGCGGCTTTGAAGGA 59.9 0.13
mm4_intron_Acad10_chr5_121656551_F TCGTCGGCAGCGTCGACCCCAATCTCCTGCCTTC 60.1 0.13
mm4_intron_Acad10_chr5_121656551_R GTCTCGTGGGCTCGGACATCTGCGGCTTTGAAGGA 59.9 0.13
mm4_intron_9530036O11Rik_chr5_28816579_F Primer3: not found at this Tm N.d. 0.12
mm4_intron_9530036O11Rik_chr5_28816579_R Primer3: not found at this Tm N.d. 0.12
mm4_exon_Pdcd7_chr9_65346298_F Primer3: not found at this Tm N.d. 0.11
mm4_exon_Pdcd7_chr9_65346298_R Primer3: not found at this Tm N.d. 0.11
mm4_exon_Arhgap1_chr2_91653205_F Primer3: not found at this Tm N.d. 0.11
mm4_exon_Arhgap1_chr2_91653205_R Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Pccb|Msl2_chr9_101053423_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Pccb|Msl2_chr9_101053423_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm25574|Ankrd50_chr3_38410198_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm25574|Ankrd50_chr3_38410198_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm12536|1700042G15Rik_chr4_57315172_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm12536|1700042G15Rik_chr4_57315172_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_6820408C15Rik|Gm14158_chr2_152450280_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_6820408C15Rik|Gm14158_chr2_152450280_R Primer3: not found at this Tm N.d. 0.10
mm4_intron_Fam114a1_chr5_65033215_F Primer3: not found at this Tm N.d. 0.10
mm4_intron_Fam114a1_chr5_65033215_R Primer3: not found at this Tm N.d. 0.10
mm4_intron_Fst_chr13_114456707_F Primer3: not found at this Tm N.d. 0.10
mm4_intron_Fst_chr13_114456707_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Ppp1r15b|Plekha6_chr1_133192458_F TCGTCGGCAGCGTCCTCTCTTATGGCTGCTCGGG 59.9 0.10
mm4_intergenic_Ppp1r15b|Plekha6_chr1_133192458_R GTCTCGTGGGCTCGGCCCAGCCCCACATGAATTGA 60.3 0.10
mm4_intergenic_Gm26419|Pak2_chr16_32079015_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm26419|Pak2_chr16_32079015_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm7308|Parp11_chr6_127425070_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Gm7308|Parp11_chr6_127425070_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Dppa3|Gm10420_chr6_122640581_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Dppa3|Gm10420_chr6_122640581_R Primer3: not found at this Tm N.d. 0.10
mm4_exon_AI480526_chr5_123141451_F Primer3: not found at this Tm N.d. 0.09
mm4_exon_AI480526_chr5_123141451_R Primer3: not found at this Tm N.d. 0.09
mm4_exon_Camk2n1_chr4_138454356_F Primer3: not found at this Tm N.d. 0.09
mm4_exon_Camk2n1_chr4_138454356_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Gm13282|Klhl9/Gm26566_chr4_88708672_F TCGTCGGCAGCGTCCAGTGTTGCTCAGGTGTTTGG 59.9 0.09
mm4_intergenic_Gm13282|Klhl9/Gm26566_chr4_88708672_R GTCTCGTGGGCTCGGAAGGATAACACTTTACAGGTGAGT 57.5 0.09
mm4_exon_Adam12_chr7_134225247_F TCGTCGGCAGCGTCGCCGTTCTCATTACCACGGA 60.1 0.09
mm4_exon_Adam12_chr7_134225247_R GTCTCGTGGGCTCGGGAGTGGATAGGACCCAGGCT 60.3 0.09
mm3_intergenic_Negr1|Gm22458_chr3_157327222_F Primer3: not found at this Tm N.d. 0.08
mm3_intergenic_Negr1|Gm22458_chr3_157327222_R Primer3: not found at this Tm N.d. 0.08
mm4_intergenic_Ap3m2|1700041G16Rik_chr8_22806747_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Ap3m2|1700041G16Rik_chr8_22806747_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Dmrt2|Gm8825_chr19_25771835_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Dmrt2|Gm8825_chr19_25771835_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_4930404H24Rik|8430427H17Rik_chr2_153528284_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_4930404H24Rik|8430427H17Rik_chr2_153528284_R Primer3: not found at this Tm N.d. 0.07
mm4_intron_Prmt2_chr10_76234140_F Primer3: not found at this Tm N.d. 0.06
mm4_intron_Prmt2_chr10_76234140_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm9901/Ccsap|Acta1_chr8_123878274_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm9901/Ccsap|Acta1_chr8_123878274_R Primer3: not found at this Tm N.d. 0.06
mm4_intron_Mrps31_chr8_22415854_F Primer3: not found at this Tm N.d. 0.06
mm4_intron_Mrps31_chr8_22415854_R Primer3: not found at this Tm N.d. 0.06
mm4_exon_Ergic3_chr2_156008080_F TCGTCGGCAGCGTCCTTTGAGGAGGCAGGTGAGG 60.0 0.06
mm4_exon_Ergic3_chr2_156008080_R GTCTCGTGGGCTCGGGGAAAGGGGACACCAGCC 59.9 0.06
mm4_intron_Mmd_chr11_90275268_F Primer3: not found at this Tm N.d. 0.06
mm4_intron_Mmd_chr11_90275268_R Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Il1rapl1|Gm25006_chrX_88192364_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Il1rapl1|Gm25006_chrX_88192364_R Primer3: not found at this Tm N.d. 0.06
mm4_intron_Fetub_chr16_22920126_F TCGTCGGCAGCGTCAGCTTTCTTGCTGCGTCATG 59.4 0.05
mm4_intron_Fetub_chr16_22920126_R GTCTCGTGGGCTCGGCCAAGGGCTCTGTTCTCAGG 60.0 0.05
mm4_intergenic_Bmpr2|Fam117b_chr1_59888998_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Bmpr2|Fam117b_chr1_59888998_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Adamts5|Gm25715_chr16_86240953_F Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Adamts5|Gm25715_chr16_86240953_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_AU022754|Mirlet7c-2_chr15_85656265_F TCGTCGGCAGCGTCGCCAGGCTCCCAACTTGTAA 60.2 0.05
mm4_intergenic_AU022754|Mirlet7c-2_chr15_85656265_R GTCTCGTGGGCTCGGCTCAGTCCGCTCCCTTTGAA 59.6 0.05
mm3_intron_Trmt1_chr8_84694688_F Primer3: not found at this Tm N.d. 0.05
mm3_intron_Trmt1_chr8_84694688_R Primer3: not found at this Tm N.d. 0.05
mm4_exon_Zfp521_chr18_13972702_F Primer3: not found at this Tm N.d. 0.05
mm4_exon_Zfp521_chr18_13972702_R Primer3: not found at this Tm N.d. 0.05
mm4_intron_Trim37_chr11_87177604_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Trim37_chr11_87177604_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Got2|Gm22780_chr8_96306036_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Got2|Gm22780_chr8_96306036_R Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_4933434M16Rik|2610035D17Rik_chr11_112941984_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_4933434M16Rik|2610035D17Rik_chr11_112941984_R Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Kat6a|Ank1_chr8_22974783_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Kat6a|Ank1_chr8_22974783_R Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Cdh11|Gm8730_chr8_102803371_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Cdh11|Gm8730_chr8_102803371_R Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Immp2l|Gm25497_chr12_41221244_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Immp2l|Gm25497_chr12_41221244_R Primer3: not found at this Tm N.d. 0.04
mm4_intron_4931406P16Rik_chr7_34261863_F TCGTCGGCAGCGTCGTGTGGGATCTGGCTGATGA 59.4 0.03
mm4_intron_4931406P16Rik_chr7_34261863_R GTCTCGTGGGCTCGGTGCCATAGGAGAAGAGGCTG 58.8 0.03
mm4_intergenic_Sec11a|Zfp592_chr7_80993452_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Sec11a|Zfp592_chr7_80993452_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Gm13318_chr2_16084400_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Gm13318_chr2_16084400_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm11445|Gm11446_chr2_159856471_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm11445|Gm11446_chr2_159856471_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Slc2a1_chr4_119109043_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Slc2a1_chr4_119109043_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Ppp1r14c|Iyd_chr10_3539440_F TCGTCGGCAGCGTCAGCCACAAGGGAAGCTTTCT 59.5 0.03
mm4_intergenic_Ppp1r14c|Iyd_chr10_3539440_R GTCTCGTGGGCTCGGCCTCGCTTGTATTGTTCTCACA 58.6 0.03
mm4_intergenic_Cd44|Pdhx_chr2_102916041_F TCGTCGGCAGCGTCAGAGCCAAGGCAGAGAGAGT 60.2 0.03
mm4_intergenic_Cd44|Pdhx_chr2_102916041_R GTCTCGTGGGCTCGGTGGGGACTTGGAGAGGCTTT 60.7 0.03
mm4_intron_Ppard_chr17_28251709_F TCGTCGGCAGCGTCGGAAGTTGTGGGTGAGGAGG 59.9 0.03
mm4_intron_Ppard_chr17_28251709_R GTCTCGTGGGCTCGGACAGCCTCTTCCCCACTAGT 59.8 0.03
mm4_intron_Pik3c2b_chr1_133054791_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Pik3c2b_chr1_133054791_R Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Nim1|Zfp131_chr13_119760793_F TCGTCGGCAGCGTCTGCAGCAATCCTGTTTTCACA 58.9 0.03
mm4_intergenic_Nim1|Zfp131_chr13_119760793_R GTCTCGTGGGCTCGGGCACCCCAACTCATCAGTCA 59.9 0.03
mm4_intergenic_Sel1l3|Smim20_chr5_53259316_F TCGTCGGCAGCGTCAGCACAAACCAGACCCCAAA 60.0 0.03
mm4_intergenic_Sel1l3|Smim20_chr5_53259316_R GTCTCGTGGGCTCGGGAGATCTTCCCTCCCCAGGT 60.0 0.03
mm3_intergenic_Gm9884|Gm24064_chr1_25971714_F Primer3: not found at this Tm N.d. 0.03
mm3_intergenic_Gm9884|Gm24064_chr1_25971714_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Mrps27_chr13_99357127_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Mrps27_chr13_99357127_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Ssbp3_chr4_106911939_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Ssbp3_chr4_106911939_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Itpr1_chr6_108213944_F TCGTCGGCAGCGTCCGGGGAAACAAGTGCTTTGG 59.9 0.02
mm4_intron_Itpr1_chr6_108213944_R GTCTCGTGGGCTCGGCGTCAGTAGCTGGCCAGAAA 60.0 0.02
mm3_intergenic_Gm22227|Zfp703_chr8_26830241_F Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_Gm22227|Zfp703_chr8_26830241_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Eno2_chr6_124766452_F TCGTCGGCAGCGTCGGCTCAGGTCTCAAGTTCCC 60.0 0.02
mm4_intron_Eno2_chr6_124766452_R GTCTCGTGGGCTCGGTGAGCACTGTACTTGCCCAG 59.9 0.02
mm4_intron_Cntnap2_chr6_46405778_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Cntnap2_chr6_46405778_R Primer3: not found at this Tm N.d. 0.02
mm4_exon_Abcb4_chr5_8893813_F Primer3: not found at this Tm N.d. 0.02
mm4_exon_Abcb4_chr5_8893813_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm26619/Gm10260|Arhgef28_chr13_97895721_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm26619/Gm10260|Arhgef28_chr13_97895721_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Fbxw10|Tvp23b_chr11_62879356_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Fbxw10|Tvp23b_chr11_62879356_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Rara_chr11_98969510_F TCGTCGGCAGCGTCTGCAGAATCGACATGAGTGATCT 59.8 0.02
mm4_intron_Rara_chr11_98969510_R GTCTCGTGGGCTCGGCAATCACTCACAGGGCCTCC 60.3 0.02
mm4_exon_Dscam_chr16_96709102_F TCGTCGGCAGCGTCGAGTGAACTTCTGGGTGGCA 59.8 0.02
mm4_exon_Dscam_chr16_96709102_R GTCTCGTGGGCTCGGACCGGATTGGCAAGAGTGAG 60.0 0.02
mm4_intron_Npas2_chr1_39330112_F TCGTCGGCAGCGTCTCTTTTCCAGTGGGCTGCTC 60.2 0.02
mm4_intron_Npas2_chr1_39330112_R GTCTCGTGGGCTCGGTCAGTCCTCATGCTCTCCCA 59.9 0.02
mm4_intergenic_Nsmce2|Trib1_chr15_59641059_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Nsmce2|Trib1_chr15_59641059_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Sppl2a_chr2_126932883_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Sppl2a_chr2_126932883_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm10030|Ltf_chr9_111015377_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm10030|Ltf_chr9_111015377_R Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_RP23-433F5.6|Cbln1_chr8_87468803_F Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_RP23-433F5.6|Cbln1_chr8_87468803_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_4930527J03Rik|B230369F24Rik_chr1_178299785_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_4930527J03Rik|B230369F24Rik_chr1_178299785_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Lphn1_chr8_83900088_F TCGTCGGCAGCGTCTAGTCCAGGCCAGGTTTTGC 60.2 0.01
mm4_exon_Lphn1_chr8_83900088_R GTCTCGTGGGCTCGGGCACCAGCAATGACAAGCC 60.0 0.01
mm4_intron_Wbscr27_chr5_134937008_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Wbscr27_chr5_134937008_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Mark3|Ckb_chr12_111666393_F Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Mark3|Ckb_chr12_111666393_R Primer3: not found at this Tm N.d. 0.01
mm4_intergenic_Defb34|Spag11b_chr8_19128040_F TCGTCGGCAGCGTCTGACTAGAGGCCCAGGAAAA 57.9 0.01
mm4_intergenic_Defb34|Spag11b_chr8_19128040_R GTCTCGTGGGCTCGGGTGCAGTAAGAGAAGGGCAGA 59.7 0.01
mm4_intergenic_Wdr66|Gm15857/Wdr66_chr5_123292265_F TCGTCGGCAGCGTCGCGGAAGGTAGAATCTGGCA 59.8 0.01
mm4_intergenic_Wdr66|Gm15857/Wdr66_chr5_123292265_R GTCTCGTGGGCTCGGGCCTTTGCTCAGGAGAGTGT 59.9 0.01
mm4_exon_Pou3f1_chr4_124660186_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Pou3f1_chr4_124660186_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm24839|Tmem132c_chr5_126756844_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm24839|Tmem132c_chr5_126756844_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Csnk2a2_chr8_95446343_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Csnk2a2_chr8_95446343_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Nrtn_chr17_56755131_F TCGTCGGCAGCGTCGGAAGGGAAACTGAGGCAGG 60.3 0.00
mm4_intron_Nrtn_chr17_56755131_R GTCTCGTGGGCTCGGGAGGCTCGGAGAACAGACAC 60.1 0.00
mm4_intron_Il17rd_chr14_27039327_F TCGTCGGCAGCGTCTCTCCGGGTTCCAGGTTACC 60.9 0.00
mm4_intron_Il17rd_chr14_27039327_R GTCTCGTGGGCTCGGGCAGGATGAGCGGCTAGAAA 60.1 0.00
mm4_intergenic_Gm5250|Prdm14_chr1_13096001_F TCGTCGGCAGCGTCTGATGCAGCTGGAAGATATGAGA 59.3 0.00
mm4_intergenic_Gm5250|Prdm14_chr1_13096001_R GTCTCGTGGGCTCGGGGGCCCTCTTTCTGCTTTTG 59.3 0.00
mm4_exon_Ptp4a2_chr4_129820547_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Ptp4a2_chr4_129820547_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.

mm0_exon_Pop5_chr5_115238018 GAGCGTGGAGATTGACAGGGCACGCTGCCTGTCCGAGCCTAGTGCCGCCTTCCTCCCGCATCCGGCGCGGACGCCATGGT
GCGGTTCAAGCACAGGTAA
mm4_intergenic_Fgfr2|Gm5903_chr7_130279582 TATGGGTCCCTCCTACAGGCCTGTGTGGAGCAGTTCCGCTCTGAGGAATCGGTCCCTGTTAGGCTGGAGGCGGGAGGAAG
AAGG
GGAAACTCAAGCGGAAAATCTTTGTTACAGCTCCTGTTTCTTCTGGGC
mm4_intron_Ptprn2_chr12_116486724 GTGGATGGTAGGCAGTGAGGGGTGAGCTGCAGCCTTGGCCCGAGCTCCCCTCAGGCTCCACACCTCCTTCCTCCCGCCTC
TGGTC
TTCAGAGGTGTTTCAACCTTGCAAAACTCAAAGAAAGGGACTGCAGAGAGGAGG
mm4_intergenic_Myh9|Txn2_chr15_77888701 ACAGAGCTCCAGGTAGAGGGATCACCATGAACAAAGGAGGCGGGAGGAAGGAGGCTGGAAGGCACATGTAGTGTGGTCCT
GGTCCTGGAGAGAAAGGGCTGATCGG
mm4_intron_Grm7_chr6_111500187 AGCCAAGATGCTGTTCCTCAGAATTTCTTCAAATTCTGGTCCTCCCACCTTCCTCCTGAATCTGGCCTCTCTCCCTGTAC
TTCCAAATGTGAACGTGTGTATATACATGGAAAACTGGAAGATGCC
mm4_intergenic_Gm10620|Cmc2_chr8_116863675 GACATCCCCTGCAGATGGAGGCCAGGCTTCGAGAAAGAGGACCAACCGTCTTCCTCCCGCCTACTGCGGTAGCCCCACGG
ACCTCTCCTTTCACCTTGGAGAAGACTAGACCCTCGCG
mm3_intergenic_Gm3839|Ptprg_chr14_11552736 TTTGCCCTCTGCCTAACCAGGCCAAGTTGCAGCCACCTTCTTCCCGAATCCGGGGGTTGGGAAAACTCCGCGACAGACCT
T
mm4_intergenic_Diap2|Gm14984_chrX_130389026 GCAACCCTACCTGTGCTCTCTATTGAATTGCACTTCCTGCTTCTGCCACCATCCTCTCACATCCGGGGCTACTACCTCTG
AAATCTTTAGATTATAAAGTCACAATCATCTAAAACCATGGCTTAGTTGTGTGAAC
mm4_intron_Maea_chr5_33365609 TGGCCTGCCAGAAGAAAACTTTTGCCACTTCTGTCAGAATAGAAGCTGTGGTAATCCCACTTCTTCCCGCACCCGGAGCT
TCAATGGTAGAAATTGTGAGACCTGTGGATCCTAGGCCAT
mm4_intergenic_Nfia|Nfia/E130114P18Rik_chr4_97775148 GCTGTAGTTGGAACTGGGCTTTGTAGTGTTCCCCCGCCTTCCACCCTCCTCCGGAGCGGGGGAGGGAGCGGGAGCTGGCA
CAGATAGGAGAGAAAGCAGGAGCGGATGTGGAGGGCTTTGGTGCTGAACTTG
mm4_intergenic_F730016J06Rik|Olfm1_chr2_28189175 ACAAGGTTAGAGGAGGGGCAGGAGTGAAACACTGGAGGCCGGAGGAAGGAGGAACTGAGCACACTAGCTGCTAGCCTT
mm4_intergenic_Gm2366/Fat1|Gm2366_chr8_45020786 AGGAGGAAAGAGAACACGTGTATAATAAAAAATAAACGTAGCCACACCGGATAGGGAAGGAAGGAGGAGTCTTCAGGCAG
GGGCATTCTGGTGAACGAA
mm4_intergenic_Foxp1|Gm24936_chr6_99522897 CTCTCAAAGCCCCTGTGACCCGCCGAGCGCGATCTGCACCCCGATGCGGCGCCCTTGGGTCGCCTGCCCCCTTCCCCCCG
CATCCCGGA
ACAAATAAATAAATAAGGCGCGCAAGAGAGG
mm4_intron_Baiap2l2_chr15_79268641 GTGGCTCTATTAAGGATGTTTCAAAGAATAGCCTCCTTCCTCCAGCAACCAGCAGGGATTGGACATCCAGACAATGGAAC
TCACTTCTGCAGGG
mm4_intron_Acad12_chr5_121614594 GACCCCAATCTCCTGCCTTCTTCTCTCACTGCAGATGTCTCTTCAGGGACCCTGGTGGCTCCCTGCTCCTTCCTCCTTCC
TCCCTCATCAGCCA
GAGAATTCCTTCAAAGCCGCAGATGT
mm4_intron_Acad10_chr5_121656551 GACCCCAATCTCCTGCCTTCTTCTCTCACTGCAGATGTCTCTTCAGGACCCTGGTGGCTCCCTGCTCCTTCCTCCTTCCT
CCCTCATCAGCCA
GAGAAATCCTTCAAAGCCGCAGATGT
mm4_intergenic_Ppp1r15b|Plekha6_chr1_133192458 CTCTCTTATGGCTGCTCGGGATGCTCAGCTGACCACCTCCTGGTGCCCAGGCCGCATGCTGGAGGAAGCAGGTGGCAAAT
TCTGTGTGTGAGGCTCACATGTTCAATTCATGTGGGGCTGGG
mm4_intergenic_Gm13282|Klhl9/Gm26566_chr4_88708672 CAGTGTTGCTCAGGTGTTTGGTTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTTACTCA
CCTGTAAAGTGTTATCCTT
mm4_exon_Adam12_chr7_134225247 GCCGTTCTCATTACCACGGAACAAATTAGCGTCTCCCCCGCACCCCGCCTTCCTGCGGCGTCCCGCGGGTCCTAGAGACC
GCTCCGGGGTCCCCGACAGCCTGGGTCCTATCCACTC
mm4_exon_Ergic3_chr2_156008080 CTTTGAGGAGGCAGGTGAGGGGGTGGAGCCCCACTTACGCACCGCTACGCCTGCGCAAGACAGCGAGGGGCGTGACCCCG
GGTCCAGGCCGGAAGCGGGAGGCTGGAGGGGCGGGCCGGCTGGTGTCCCCTTTCC
mm4_intron_Fetub_chr16_22920126 AGCTTTCTTGCTGCGTCATGCGTGGGCGGGAGGCGGGAGGCAGGAGGAAGAACTCTGCCTAACAAATGTACTCCCCTGAC
ATGGGCATGCCCCACCCCTGAGAACAGAGCCCTTGG
mm4_intergenic_AU022754|Mirlet7c-2_chr15_85656265 GCCAGGCTCCCAACTTGTAAGATCCCACCCGAATTGGAAAGAAGGTGTCCTCCAAGGATGGGGGAGGAAGGCAGTCAGAG
CTGTGAAGGCTGTAGAAGTCACCCTGGAGTGGGTGTATGTTTCAAAGGGAGCGGACTGAG
mm4_intron_4931406P16Rik_chr7_34261863 GTGTGGGATCTGGCTGATGAAGAATGCAATGCTACTGGCAACTGCAGCGGCTCTGGAAACTGCCTCCTGCCTCCTGCCTC
AGGCG
GAACACTGATGAGACTATGTAGTTGAGTCTTCTCAGCCTCTTCTCCTATGGCA
mm4_intergenic_Ppp1r14c|Iyd_chr10_3539440 AGCCACAAGGGAAGCTTTCTGTGAGTAATGGTTCTCTCTCCCATCCTTCCTTCCTCCCTCATCCCCAGCTGTGAGAACAA
TACAAGCGAGG
mm4_intergenic_Cd44|Pdhx_chr2_102916041 AGAGCCAAGGCAGAGAGAGTTCTGAGAGGCTGGAGGTGGGAGGAAGGAGAAAAGCCTCTCCAAGTCCCCA
mm4_intron_Ppard_chr17_28251709 GGAAGTTGTGGGTGAGGAGGTCCTGGTCCTTCTCTGGATGCTGGAGGGAGGTGGGGGAGGCTGGCTTTTAAAGGTCCCTC
CAACAAAGACAGCACTAGTGGGGAAGAGGCTGT
mm4_intergenic_Nim1|Zfp131_chr13_119760793 TGCAGCAATCCTGTTTTCACAAAGTTCTCCCTTGTCTCTCTTTTGACTCTCCCTTCTCCTTCCTCCCTCTTCCAGTGATA
CCTGGAATGCTTTTCACATAATGTTATTTCTGACTGATGAGTTGGGGTGC
mm4_intergenic_Sel1l3|Smim20_chr5_53259316 AGCACAAACCAGACCCCAAAGAACCTCCGGGTGGGGGAGGGAGGGGGAGGGAGGGGCCCACCTGGGGAGGGAAGATCTC
mm4_intron_Itpr1_chr6_108213944 CGGGGAAACAAGTGCTTTGGGATCTCAACACGTCCCTTACTTCCATCCACCTTCAATCCCCATCCGGCGTGTTCGAAGTT
TCTGCGTGTCTGATCTTTCTGGCCAGCTACTGACG
mm4_intron_Eno2_chr6_124766452 GGCTCAGGTCTCAAGTTCCCATCCTTTCACCTTAGCTGCCTGTTCACCTTCCTCCCTCATCCCTCACCTGGTTGCATGCT
CCCTCCTCCCCTGGGAGAGGCTGGGCAAGTACAGTGCTCA
mm4_intron_Rara_chr11_98969510 TGCAGAATCGACATGAGTGATCTCCAAATTATGCCAGCTTNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNGCGTCAGCAGCCGCCACGGAGGCCCTGTGAGTGATTG
mm4_exon_Dscam_chr16_96709102 GAGTGAACTTCTGGGTGGCAGGCTCTAACCGGTGCCACCTACCTGCCTCATCCGCCGTGATGGTGATCTCGTTGCTGGGC
TCACTCTTGCCAATCCGGT
mm4_intron_Npas2_chr1_39330112 TCTTTTCCAGTGGGCTGCTCAGTCTCTCAGAGAGGATGCATGACACCCTTGCTCTCTCCTTCCTCCCGCATCGTAGGAAG
AAGCTAGGTCTGGGAGAGCATGAGGACTGA
mm4_exon_Lphn1_chr8_83900088 TAGTCCAGGCCAGGTTTTGCGCTTGTCCGCCGCCCCCCTCCCGGAGCCGGCGGGCCCGGCAGCGCCGAGAGCAGGCGGCT
TGTCATTGCTGGTGC
mm4_intergenic_Defb34|Spag11b_chr8_19128040 TGACTAGAGGCCCAGGAAAAAGCTTCTCTAATTCCTTGTCAATTTTAACACCAGTNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNGATCTGCCCTTCTCTTACTGCAC
mm4_intergenic_Wdr66|Gm15857/Wdr66_chr5_123292265 GCGGAAGGTAGAATCTGGCACAGTGTTTAAAATCCCTGAGAACACATGGAGCTTCAGGAGGCTCTCCTTCCTCCCTCTTC
AGGGG
ACACACTCTCCTGAGCAAAGGC
mm4_intron_Nrtn_chr17_56755131 GGAAGGGAAACTGAGGCAGGCGCCGGATGCGGCAGTAACAGGGTGGGGAGGGAGGGCACTTGCCTGTCCTCTCCTTCCTT
CCCAAATTCAAAGTTTGGTCTCAGGTCCTAGATAAAGGTGTCTGTTCTCCGAGCCTC
mm4_intron_Il17rd_chr14_27039327 TCTCCGGGTTCCAGGTTACCTGGGGGCGCGCCGCGCGCACAGGTGTTCGCGCGCAGGCGAGCGGGGACCGCGGGGATCGC
TGGACGCCGGATCCGCGAGCAACGGGGTTCTCAGCTTTTCTAGCCGCTCATCCTGC
mm4_intergenic_Gm5250|Prdm14_chr1_13096001 TGATGCAGCTGGAAGATATGAGANNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNTCAAAAGCAGAAAGAGGGCCC

Input file for Crispresso

Crispresso, written by Luca Pinello, is a software package to quantify the Cas9-induced mutations on off- or on-targets.

Click here to download an amplicon input file for Crispresso. For each off-target, it includes the off-target name, its PCR amplicon and the guide sequence. Keep a copy of this file.

After sequencing, run CRISPRessoPooled. The tool will map the reads to the amplicons and analyse the mutations:
CRISPRessoPooled -r1 Reads1.fastq.gz -r2 Reads2.fastq.gz -f crisporAmplicons_ozJbN0W08qlgEBn3RvZK.txt --name MY_EXPERIMENT

Version 5.01 - Documentation  - Contact us - Downloads/local installation - Citation - License
CRISPR/Cas9 Guide Designer for chordate vertebrate ecdysozoans lophotrochozoans protostomes spongi corals plants butterflies metazoans genomes fruitflies insects nematodes mammals.