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Contents:

PCR primers for off-targets of CCAGAGAGGCTGTCCGCGGA GGG

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

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

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

Maximum amplicon length:     Primer Tm:

NamePrimer Sequence Tm CFD Score
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52314138_F TCGTCGGCAGCGTCGTCGATGGACTAGCAGGCAG 60.2 1.00
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52314138_R GTCTCGTGGGCTCGGAGCTACTAGGTTGCCCTTGG 59.0 1.00
mm4_intergenic_Asic2|4930527B05Rik_chr11_81316396_F TCGTCGGCAGCGTCGGACCAATGGCACTGTCTCA 59.9 0.33
mm4_intergenic_Asic2|4930527B05Rik_chr11_81316396_R GTCTCGTGGGCTCGGACACTGAACATGAAGGGCTGT 59.8 0.33
mm4_intron_Kctd10_chr5_114376800_F TCGTCGGCAGCGTCGCAGTCATGACCAAACACATCC 60.0 0.30
mm4_intron_Kctd10_chr5_114376800_R GTCTCGTGGGCTCGGCCACCACACACAAACACACC 59.8 0.30
mm4_intergenic_Pitpnb|Mn1_chr5_111398283_F TCGTCGGCAGCGTCTTGTGTGGAGCTCCGTGTAC 59.9 0.23
mm4_intergenic_Pitpnb|Mn1_chr5_111398283_R GTCTCGTGGGCTCGGCAACATCTCCCTCTCGTGCA 59.7 0.23
mm4_exon_Nacc2/Gm13554_chr2_26060073_F TCGTCGGCAGCGTCCTTCTTGTTGCTGTGGCCAC 59.9 0.23
mm4_exon_Nacc2/Gm13554_chr2_26060073_R GTCTCGTGGGCTCGGGGGTGGCTCTGTTATCCAGG 59.8 0.23
mm4_intergenic_Mcm5|Rasd2_chr8_75162825_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Mcm5|Rasd2_chr8_75162825_R Primer3: not found at this Tm N.d. 0.22
mm4_intron_Themis2_chr4_132792839_F TCGTCGGCAGCGTCCCCCTAGTGTAGCCCTCAGG 60.7 0.20
mm4_intron_Themis2_chr4_132792839_R GTCTCGTGGGCTCGGAGAGCCAGGAGAAAGTGCAC 59.9 0.20
mm4_intergenic_Tango6|Has3_chr8_106858924_F Primer3: not found at this Tm N.d. 0.19
mm4_intergenic_Tango6|Has3_chr8_106858924_R Primer3: not found at this Tm N.d. 0.19
mm4_intron_Prkce_chr17_86418257_F TCGTCGGCAGCGTCCAAGGGTTGGGCCTCTATGG 60.1 0.18
mm4_intron_Prkce_chr17_86418257_R GTCTCGTGGGCTCGGTCTTACCCAGCTTGCACCTG 59.9 0.18
mm4_intron_Lmx1a_chr1_167789815_F TCGTCGGCAGCGTCGGCCATACAGCACACTCCTT 60.0 0.18
mm4_intron_Lmx1a_chr1_167789815_R GTCTCGTGGGCTCGGAGCTTTGAGTGGAGGAAGCC 59.9 0.18
mm3_intergenic_Smlr1|Tmem200a_chr10_25696510_F Primer3: not found at this Tm N.d. 0.17
mm3_intergenic_Smlr1|Tmem200a_chr10_25696510_R Primer3: not found at this Tm N.d. 0.17
mm4_intron_Fryl_chr5_73052957_F TCGTCGGCAGCGTCCAGTGGATGAGCTCTGACGG 60.1 0.16
mm4_intron_Fryl_chr5_73052957_R GTCTCGTGGGCTCGGTCTGTGATCTCATGCACCCC 59.4 0.16
mm4_intergenic_Gm833|Gm25779_chr4_152790533_F TCGTCGGCAGCGTCCAGGAGAGGAACACCTGCTC 59.7 0.14
mm4_intergenic_Gm833|Gm25779_chr4_152790533_R GTCTCGTGGGCTCGGTCTTCCCGCGTAGACTGAGA 60.0 0.14
mm4_intron_Nfyb_chr10_82758839_F TCGTCGGCAGCGTCGCGTTCACTTGTCTGAAGCC 59.7 0.10
mm4_intron_Nfyb_chr10_82758839_R GTCTCGTGGGCTCGGCCCATGGAGCAGTCACTAGC 60.1 0.10
mm4_intron_Rad50_chr11_53704584_F Primer3: not found at this Tm N.d. 0.10
mm4_intron_Rad50_chr11_53704584_R Primer3: not found at this Tm N.d. 0.10
mm4_exon_Scn3b_chr9_40269328_F TCGTCGGCAGCGTCTTGATCACCCAATCGCCCAG 60.3 0.10
mm4_exon_Scn3b_chr9_40269328_R GTCTCGTGGGCTCGGGACTGGCGATCTCAGAGGTG 59.8 0.10
mm4_intergenic_Osbpl3|Gm26289_chr6_50457480_F TCGTCGGCAGCGTCTCTCTGAGGCTCTTTCTCACA 58.1 0.09
mm4_intergenic_Osbpl3|Gm26289_chr6_50457480_R GTCTCGTGGGCTCGGTCTTCGTCAAACTTCCCCGG 59.9 0.09
mm4_intergenic_1700041M05Rik|Grik3_chr4_125396722_F TCGTCGGCAGCGTCTGGCATATCACCCCAAAGCA 59.6 0.08
mm4_intergenic_1700041M05Rik|Grik3_chr4_125396722_R GTCTCGTGGGCTCGGGAGCTGTCCAGAAGGCACAT 60.0 0.08
mm4_intergenic_Klf11|Cys1_chr12_24665396_F TCGTCGGCAGCGTCATACCTGGTACCTGGGCAGT 59.9 0.07
mm4_intergenic_Klf11|Cys1_chr12_24665396_R GTCTCGTGGGCTCGGGCCGACTGATGAGCTGTGAT 60.1 0.07
mm4_intergenic_Pars2|Ttc4_chr4_106658461_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Pars2|Ttc4_chr4_106658461_R Primer3: not found at this Tm N.d. 0.07
mm4_exon_Micall1_chr15_79120773_F TCGTCGGCAGCGTCCGGTAACTGTACCCCGTTCC 60.1 0.07
mm4_exon_Micall1_chr15_79120773_R GTCTCGTGGGCTCGGCCAAGTACCGCTGCACCA 59.9 0.07
mm4_intron_Ttc3_chr16_94400311_F Primer3: not found at this Tm N.d. 0.07
mm4_intron_Ttc3_chr16_94400311_R Primer3: not found at this Tm N.d. 0.07
mm4_intron_Ccdc85c_chr12_108208831_F TCGTCGGCAGCGTCGGTTGTCCTCAGTTAGGCCC 60.0 0.06
mm4_intron_Ccdc85c_chr12_108208831_R GTCTCGTGGGCTCGGAGATCGCTCCATGCCTATGC 60.0 0.06
mm4_intergenic_Gm13090|Camta1_chr4_151102334_F Primer3: not found at this Tm N.d. 0.06
mm4_intergenic_Gm13090|Camta1_chr4_151102334_R Primer3: not found at this Tm N.d. 0.06
mm4_intron_Mxi1_chr19_53348159_F TCGTCGGCAGCGTCAATCTGGTGTCCCTCTGTGC 59.6 0.05
mm4_intron_Mxi1_chr19_53348159_R GTCTCGTGGGCTCGGGAAGAGAGCTGTGAGGCCTG 60.1 0.05
mm4_intergenic_Rnf130|Mir340_chr11_50058302_F TCGTCGGCAGCGTCAGTGGCTTTAAGTCTGGGCT 58.9 0.05
mm4_intergenic_Rnf130|Mir340_chr11_50058302_R GTCTCGTGGGCTCGGGCCTGTAATTACCTCGCCCA 59.8 0.05
mm4_intron_Rad50_chr11_53662977_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Rad50_chr11_53662977_R Primer3: not found at this Tm N.d. 0.05
mm4_intron_Lmtk3_chr7_45785255_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Lmtk3_chr7_45785255_R Primer3: not found at this Tm N.d. 0.05
mm4_intergenic_Mrpl36|Lpcat1_chr13_73416792_F TCGTCGGCAGCGTCAAAGAGGTGGATGTCCGCTG 60.0 0.04
mm4_intergenic_Mrpl36|Lpcat1_chr13_73416792_R GTCTCGTGGGCTCGGCACCTCACCTGCCATTAGCT 59.7 0.04
mm4_exon_Stk11ip_chr1_75529128_F TCGTCGGCAGCGTCTCCAGCCATAGAGGACGACA 60.0 0.04
mm4_exon_Stk11ip_chr1_75529128_R GTCTCGTGGGCTCGGGGCTCTGGCTCCACCCTG 62.1 0.04
mm4_intergenic_Wdr20a|Stk30_chr12_110797404_F TCGTCGGCAGCGTCCACTGGACAGTTGGACCCTC 59.9 0.03
mm4_intergenic_Wdr20a|Stk30_chr12_110797404_R GTCTCGTGGGCTCGGCTACAGGGCAGCTGCTTTCT 60.0 0.03
mm4_intron_Rps6kc1_chr1_190723236_F TCGTCGGCAGCGTCGGGAGGGAAGAGCCATGAAA 59.3 0.03
mm4_intron_Rps6kc1_chr1_190723236_R GTCTCGTGGGCTCGGGGGTGGATGAGCTCATGACC 60.1 0.03
mm4_intron_Ppp1r16a_chr15_76671926_F TCGTCGGCAGCGTCGTGAGGGGCCTGCAGAAG 60.0 0.03
mm4_intron_Ppp1r16a_chr15_76671926_R GTCTCGTGGGCTCGGATGGATTTGCCCGACCAACA 60.2 0.03
mm4_intron_Fkbp6_chr5_135348157_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Fkbp6_chr5_135348157_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Pacsin2_chr15_83443647_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Pacsin2_chr15_83443647_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Kap|Etv6_chr6_133902220_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Kap|Etv6_chr6_133902220_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm10709|Tmem123_chr9_7752666_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm10709|Tmem123_chr9_7752666_R Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Coq3|Faxc_chr4_21923663_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Coq3|Faxc_chr4_21923663_R Primer3: not found at this Tm N.d. 0.02
mm4_intron_Prrc1_chr18_57356240_F Primer3: not found at this Tm N.d. 0.02
mm4_intron_Prrc1_chr18_57356240_R Primer3: not found at this Tm N.d. 0.02
mm3_intergenic_Spata2|Rnf114/Spata2_chr2_167492119_F TCGTCGGCAGCGTCGGGCCTTCCAAAGCTCTGAA 60.2 0.02
mm3_intergenic_Spata2|Rnf114/Spata2_chr2_167492119_R GTCTCGTGGGCTCGGTAGAGAGGGGACTTGGCTCC 60.0 0.02
mm4_intergenic_Mtx3|Cmya5_chr13_92987491_F TCGTCGGCAGCGTCTGTGTGGCTCTTTCCTGTCC 59.8 0.02
mm4_intergenic_Mtx3|Cmya5_chr13_92987491_R GTCTCGTGGGCTCGGTAAGCATCACGGAGGGGATG 59.2 0.02
mm4_intron_Phf17_chr3_41584370_F TCGTCGGCAGCGTCCAGAAGGTGGAGTGAGTTGGA 59.3 0.02
mm4_intron_Phf17_chr3_41584370_R GTCTCGTGGGCTCGGATGACAACCCGGTGTGTACC 59.9 0.02
mm4_intergenic_Ilkap|Hes6_chr1_91406008_F TCGTCGGCAGCGTCAGCAAACCTGATCCCTCTGC 60.0 0.01
mm4_intergenic_Ilkap|Hes6_chr1_91406008_R GTCTCGTGGGCTCGGCTCCCAGAAACCTGTGCGT 59.9 0.01
mm4_intron_Prkcd_chr14_30599538_F Primer3: not found at this Tm N.d. 0.01
mm4_intron_Prkcd_chr14_30599538_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Pcdh15_chr10_74631591_F Primer3: not found at this Tm N.d. 0.01
mm4_exon_Pcdh15_chr10_74631591_R Primer3: not found at this Tm N.d. 0.01
mm4_exon_Rusc1_chr3_89091214_F TCGTCGGCAGCGTCGCTTCTTACCCACGGCGT 60.0 0.01
mm4_exon_Rusc1_chr3_89091214_R GTCTCGTGGGCTCGGAAAATCGACTTGGGCTGCAG 59.1 0.01
mm4_exon_Krt17_chr11_100257640_F TCGTCGGCAGCGTCCGGTTCTCTGTCTCTGCCAG 60.1 0.01
mm4_exon_Krt17_chr11_100257640_R GTCTCGTGGGCTCGGGGAGTCAGCTTTGCCACTCT 59.9 0.01
mm4_exon_Rnf150_chr8_82863914_F TCGTCGGCAGCGTCCAGGGAGTGAGTCGCAGC 60.1 0.01
mm4_exon_Rnf150_chr8_82863914_R GTCTCGTGGGCTCGGTGTCATCTCTCTCCTCCGGG 60.1 0.01
mm4_intergenic_Parva|2310014F06Rik_chr7_112597411_F TCGTCGGCAGCGTCCTGCCATTTCAGGGTCGGAA 60.3 0.01
mm4_intergenic_Parva|2310014F06Rik_chr7_112597411_R GTCTCGTGGGCTCGGGGTCTGGCCTGAGAAACGAA 59.9 0.01
mm4_intron_Flt1_chr5_147725260_F TCGTCGGCAGCGTCCCTCCTTTGCTCTGGTGGAG 60.0 0.01
mm4_intron_Flt1_chr5_147725260_R GTCTCGTGGGCTCGGCCTCAAGGGCTAGTTGGAGC 60.1 0.01
mm4_intergenic_Gm12066|Gm12068_chr11_24373508_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm12066|Gm12068_chr11_24373508_R Primer3: not found at this Tm N.d. 0.00
mm3_exon_Tssc4/Trpm5_chr7_143070091_F Primer3: not found at this Tm N.d. 0.00
mm3_exon_Tssc4/Trpm5_chr7_143070091_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Rapgef1_chr2_29706221_F TCGTCGGCAGCGTCAGCTAATGTGCGATGTGGGT 59.7 0.00
mm4_intron_Rapgef1_chr2_29706221_R GTCTCGTGGGCTCGGGGAAGGCTGATTCTCTGCGT 60.1 0.00
mm4_intron_Masp1_chr16_23496292_F TCGTCGGCAGCGTCCCAGCACCTGGTCTTCTGTT 59.8 0.00
mm4_intron_Masp1_chr16_23496292_R GTCTCGTGGGCTCGGATGCTGCCTCATCTGCTTGT 60.0 0.00
mm4_intron_Cdc42bpb_chr12_111330554_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Cdc42bpb_chr12_111330554_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm26844|Gm4812_chr13_69316067_F TCGTCGGCAGCGTCAGTGGCTGAGCTGTGATGAC 60.0 0.00
mm4_intergenic_Gm26844|Gm4812_chr13_69316067_R GTCTCGTGGGCTCGGACAACTTGTGCCTACAGGAGA 58.9 0.00
mm4_exon_Ccdc62_chr5_123954507_F TCGTCGGCAGCGTCCAACGTCTACCTGGGCCTTT 59.9 0.00
mm4_exon_Ccdc62_chr5_123954507_R GTCTCGTGGGCTCGGCTCAGATGTTCCTGTGCGGT 60.0 0.00
mm4_exon_1700025G04Rik_chr1_151885856_F TCGTCGGCAGCGTCAAGGGCTGCCTGGAATTCTC 60.0 0.00
mm4_exon_1700025G04Rik_chr1_151885856_R GTCTCGTGGGCTCGGGGCAGAGATTCACATGGGGT 59.7 0.00
mm4_intron_Zfp536_chr7_37519268_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Zfp536_chr7_37519268_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Vps8_chr16_21430895_F TCGTCGGCAGCGTCCAAGTCCCTTTGTGCTTGGC 59.9 0.00
mm4_intron_Vps8_chr16_21430895_R GTCTCGTGGGCTCGGGTGGAACAGTGTGAACATCTGTG 59.9 0.00
mm4_intron_Trak1_chr9_121371653_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Trak1_chr9_121371653_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tmem9_chr1_136025743_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tmem9_chr1_136025743_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Tgfbr3_chr5_107120114_F TCGTCGGCAGCGTCGAGGCAGGAGGTGACACAAG 60.3 0.00
mm4_intron_Tgfbr3_chr5_107120114_R GTCTCGTGGGCTCGGTAAGCTTCGTGGCCTGAAGG 60.0 0.00
mm4_intron_Socs3_chr11_117969289_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Socs3_chr11_117969289_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Sh3pxd2a_chr19_47342761_F TCGTCGGCAGCGTCCCCGCACCCAGAAAGAATCT 60.0 0.00
mm4_intron_Sh3pxd2a_chr19_47342761_R GTCTCGTGGGCTCGGCCCCAAGAGCAAGAGTGAGG 60.0 0.00
mm4_intron_Rwdd2a_chr9_86572362_F TCGTCGGCAGCGTCCTATGCACCTGCGCGAGAT 60.3 0.00
mm4_intron_Rwdd2a_chr9_86572362_R GTCTCGTGGGCTCGGGCTAGGAAGGATGCCTGCAG 60.5 0.00
mm4_intron_Mfsd12_chr10_81360837_F TCGTCGGCAGCGTCCCCTGGTGTACCCTAAAGCC 59.7 0.00
mm4_intron_Mfsd12_chr10_81360837_R GTCTCGTGGGCTCGGGAGACAGGCTACACACCCAC 60.0 0.00
mm4_intron_Map2k5_chr9_63168165_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Map2k5_chr9_63168165_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Klhl22_chr16_17783996_F TCGTCGGCAGCGTCTAGAAACACCTGTGGGCAGC 60.2 0.00
mm4_intron_Klhl22_chr16_17783996_R GTCTCGTGGGCTCGGCTCACAAGGTGGCCTGGAAA 60.1 0.00
mm4_intron_Col14a1_chr15_55429686_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Col14a1_chr15_55429686_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Cntn1_chr15_92088876_F TCGTCGGCAGCGTCGCCATTTAACCCTGCATGCC 60.1 0.00
mm4_intron_Cntn1_chr15_92088876_R GTCTCGTGGGCTCGGTCATTGGTGGTGAAGAAACAACA 58.9 0.00
mm4_intron_Cdh13_chr8_119137573_F Primer3: not found at this Tm N.d. 0.00
mm4_intron_Cdh13_chr8_119137573_R Primer3: not found at this Tm N.d. 0.00
mm4_intron_Arhgap26_chr18_38691553_F TCGTCGGCAGCGTCTAGAGGCCTGATCCGTTCCA 60.0 0.00
mm4_intron_Arhgap26_chr18_38691553_R GTCTCGTGGGCTCGGTGGTTCCTACAGACTTGGCC 59.3 0.00
mm4_intron_Ankrd35_chr3_96689661_F TCGTCGGCAGCGTCGCGGTCTCGGGTGTTCATTA 60.1 0.00
mm4_intron_Ankrd35_chr3_96689661_R GTCTCGTGGGCTCGGGGGAGCCAGAAAGAGAAGCA 59.6 0.00
mm4_intron_Akap2_chr4_57808029_F TCGTCGGCAGCGTCCCCCTCTTGACAGGTAGGGA 59.9 0.00
mm4_intron_Akap2_chr4_57808029_R GTCTCGTGGGCTCGGCACCCCAAGTGTCAGAGACC 59.9 0.00
mm4_intergenic_Pla2g2e|Otud3_chr4_138893379_F TCGTCGGCAGCGTCGCCACCTTGTGAGCTACCAT 60.0 0.00
mm4_intergenic_Pla2g2e|Otud3_chr4_138893379_R GTCTCGTGGGCTCGGTTGGCACTGTTCCTCCAGTC 59.8 0.00
mm4_intergenic_Pard3|mmu-mir-21c_chr8_128054805_F TCGTCGGCAGCGTCGCACACAAATGCAGGGTCAG 60.0 0.00
mm4_intergenic_Pard3|mmu-mir-21c_chr8_128054805_R GTCTCGTGGGCTCGGTGAGACGGAACATCAACCACA 59.5 0.00
mm4_intergenic_Igkv4-86|Igkv13-84_chr6_68917551_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Igkv4-86|Igkv13-84_chr6_68917551_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Hmga2|Gm24298_chr10_120477282_F TCGTCGGCAGCGTCGCCAGCAGTGGGGTTAAAA 58.2 0.00
mm4_intergenic_Hmga2|Gm24298_chr10_120477282_R GTCTCGTGGGCTCGGAAGCTCTCCAGGGTGACTCT 59.8 0.00
mm4_intergenic_Gm24737|Lhfpl2_chr13_93984681_F TCGTCGGCAGCGTCCATCGAAGGATGGCCCAAGG 60.8 0.00
mm4_intergenic_Gm24737|Lhfpl2_chr13_93984681_R GTCTCGTGGGCTCGGGCTGAGTTCACATTTGCCCC 59.7 0.00
mm4_intergenic_Gm23847|Fgfr2_chr7_130150008_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm23847|Fgfr2_chr7_130150008_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm23120|5S_rRNA_chr11_107851510_F TCGTCGGCAGCGTCGACTTCCCAAGCCTCTGACC 60.0 0.00
mm4_intergenic_Gm23120|5S_rRNA_chr11_107851510_R GTCTCGTGGGCTCGGAAGTCTGGACCAAAGCGAGG 59.9 0.00
mm4_intergenic_Gm22670|Gm12434_chr4_47953890_F TCGTCGGCAGCGTCTCTGCCCTGATGCTCTACCT 60.0 0.00
mm4_intergenic_Gm22670|Gm12434_chr4_47953890_R GTCTCGTGGGCTCGGATTGAGGGCATTGGCTTGTT 58.3 0.00
mm4_intergenic_Gm20672|Abcg2_chr6_58545186_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm20672|Abcg2_chr6_58545186_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gm14491|Gm14512_chrX_11956264_F TCGTCGGCAGCGTCCTTTCTGTGGCACATGCAGG 59.7 0.00
mm4_intergenic_Gm14491|Gm14512_chrX_11956264_R GTCTCGTGGGCTCGGTCCTCCAGGAAGGCCTTTGA 60.4 0.00
mm4_intergenic_Gdf15|Pgpep1_chr8_70637652_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Gdf15|Pgpep1_chr8_70637652_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_Akt2-ps|Gm12663_chr11_16618265_F TCGTCGGCAGCGTCTAGACACTGGTGGGCTCAGA 59.8 0.00
mm4_intergenic_Akt2-ps|Gm12663_chr11_16618265_R GTCTCGTGGGCTCGGGCACAGATCCCTCTCAGTGG 59.8 0.00
mm4_intergenic_2610037D02Rik|4833422M21Rik_chr15_96218327_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_2610037D02Rik|4833422M21Rik_chr15_96218327_R Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_2410088K16Rik|Sh3bp4_chr1_88879641_F TCGTCGGCAGCGTCTGCAGATACAAGCGATGGCA 60.1 0.00
mm4_intergenic_2410088K16Rik|Sh3bp4_chr1_88879641_R GTCTCGTGGGCTCGGTGCTGAGGCCTCAACAAACA 60.1 0.00
mm4_exon_Lrrc24_chr15_76718359_F TCGTCGGCAGCGTCCTGGACCAAGCCCACAAATG 59.3 0.00
mm4_exon_Lrrc24_chr15_76718359_R GTCTCGTGGGCTCGGCACAACAATACCCTGCGTGC 60.1 0.00
mm4_exon_Lbh_chr17_72918359_F TCGTCGGCAGCGTCGGAGAGCTTGGTCTGCACTG 60.6 0.00
mm4_exon_Lbh_chr17_72918359_R GTCTCGTGGGCTCGGTGAATGAGATTGGCAGGGGC 60.3 0.00
mm3_intron_Drosha_chr15_12853326_F TCGTCGGCAGCGTCATTGGTCTGGTGCTCCTGTG 59.9 0.00
mm3_intron_Drosha_chr15_12853326_R GTCTCGTGGGCTCGGTGACTTCGTCATCAGCACGG 60.3 0.00
mm3_intergenic_Fam25c|Sncg_chr14_34357763_F TCGTCGGCAGCGTCCAAAAGTCAGCCCACACAGC 59.9 0.00
mm3_intergenic_Fam25c|Sncg_chr14_34357763_R GTCTCGTGGGCTCGGCTGGGCAAAGGGTAGTCGAG 60.1 0.00

Off-target amplicon sequences with primers

These only list off-targets that have primers in the table above. Primers underlined, off-targets in bold.

ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52314138 GTCGATGGACTAGCAGGCAGTGCTGCAGGGCTGGGCGCCGAGCCACGGTCTGCTGGAGCGGCCATGCTTGGCCCGGGACC
CCCAGTCCCCTCCGCGGACAGCCTCTCTGGCCAAGGGCAACCTAGTAGCT
mm4_intergenic_Asic2|4930527B05Rik_chr11_81316396 GGACCAATGGCACTGTCTCAAGTGGGTTGGCCCCATCCTTGGACAGCTTCCCTGGTGCCAGTAGAAATGACTGGCATTTT
CTAATTGGTCAACTTAAACAGCCCTTCATGTTCAGTGT
mm4_intron_Kctd10_chr5_114376800 GCAGTCATGACCAAACACATCCTCCAGAGATACTGTCCACGGGTGGCAGGATATTCTAGGAAGAAAGCATTTCCCAAAAC
TTTGATGACCTGGGGGCAAAAGAAAGGCTCACTGGTGTGTTTGTGTGTGGTGG
mm4_intergenic_Pitpnb|Mn1_chr5_111398283 TTGTGTGGAGCTCCGTGTACTGGGTTTTTTCTGTTCTCATTCTGGCATCCCTTCCTGAGGCCCTTTCCATCCACGGGCAG
CCCCTCTGC
GTCTTTCCTGATCCCTCTGCCAGCGCTGCACGAGAGGGAGATGTTG
mm4_exon_Nacc2/Gm13554_chr2_26060073 CTTCTTGTTGCTGTGGCCACTGGTGTCTGAGGCCTACTGGAGCTGGTGTTGCCCTGCTCAAAGGCCTGAGGGGAGCTCTG
GCCATCAGCGGGCAGCTGCTCTGGGGCAGCCACCTCCTGGATAACAGAGCCACCC
mm4_intron_Themis2_chr4_132792839 CCCCTAGTGTAGCCCTCAGGCTTCTTCCTTACTTCTCCCTGCTAAAGGGCAGCTGGCTCAGCCGGCCATCCTCGGACAGC
TTCCCCGG
TCTTCACTGACCACCACAGGCCTGATGGTGCACTTTCTCCTGGCTCT
mm4_intron_Prkce_chr17_86418257 CAAGGGTTGGGCCTCTATGGCTAATCAGTAATAGGCTGTGGCTATCCATCTCCAGGGAAGGTGTCCGCGTAGGGACAGGG
GATGCACCACAGGTGCAAGCTGGGTAAGA
mm4_intron_Lmx1a_chr1_167789815 GGCCATACAGCACACTCCTTCCAATCCTGCAGATCGCATCCTGTCCTGGGAGGCTGTCCGTGGGTGGCTCCAGGATCCAT
GGCAGATGACCTTGTTAATCTTTTTCAAGCTTTGGCTTCCTCCACTCAAAGCT
mm4_intron_Fryl_chr5_73052957 CAGTGGATGAGCTCTGACGGGCTAAGAGCATCAGGAAAAGCAACTTCTCCCCTCCGCGGAGAGCACCCCTGGAACTAATC
TATACTTGCCCAAAAACATTGCTATGGGGGTGCATGAGATCACAGA
mm4_intergenic_Gm833|Gm25779_chr4_152790533 CAGGAGAGGAACACCTGCTCTTCATGACCTGAGCTCCCTGCTGCAGGAAGACCTTCCACGGACAGCCACTCAGACTCAGT
CTCAGTCTACGCGGGAAGA
mm4_intron_Nfyb_chr10_82758839 GCGTTCACTTGTCTGAAGCCAACAGCAGCAGCAACACTCCTTCAGCTGACAGCCTCTCTTAGTGTGTGATGACTGTGTCT
CTCATGTGCTAGTGACTGCTCCATGGG
mm4_exon_Scn3b_chr9_40269328 TTGATCACCCAATCGCCCAGCATCGGGGCTTCGCAGGATCCAGAGACGCGCCCCGCGGAAGGGGGTCCCCCCCTCTACCC
AGCCTCCACCTCTGAGATCGCCAGTC
mm4_intergenic_Osbpl3|Gm26289_chr6_50457480 TCTCTGAGGCTCTTTCTCACATCTGTATAAACCATGTTTCATTCACTAGGTTTGGATATTCACTTTCTGGAACCTTCCAC
GGATACACTCTCTGG
AACTGTCTTTGGCTCTGCCAGGTGGCCGGGGAAGTTTGACGAAGA
mm4_intergenic_1700041M05Rik|Grik3_chr4_125396722 TGGCATATCACCCCAAAGCATCTTGCTTTCACACATGCCTGTTGGCTTTAGCAAGGGGCTTCGTGACCCCAGAGAAGCTC
TCCGCTGCTGG
CCCCCCAGGTGGCCCTTCTGCATGTGCCTTCTGGACAGCTC
mm4_intergenic_Klf11|Cys1_chr12_24665396 ATACCTGGTACCTGGGCAGTACAGGAGATGCTCCTGTAGGCTGCAGTGATACCCAAGCCCACCTCGTGCAGCCTCTCTGG
AGCCTGAGTTTCCCAGGATCACAGCTCATCAGTCGGC
mm4_exon_Micall1_chr15_79120773 CGGTAACTGTACCCCGTTCCCTGCAGGGTGACGACCTCTCCCCAGACAGCCTGTCCGAGCAGGGCAAACAGCAGCCCCCG
AGCAGCGCATGCGCAGCCTGTGGGCAGCGGGTGCACCTGGTGCAGCGGTACTTGG
mm4_intron_Ccdc85c_chr12_108208831 GGTTGTCCTCAGTTAGGCCCAGGGCCTTCCACTGACAGCCTGTCTGCCAAGCCCGTTTCTCTCAGGCCATCTGGCAAGGA
AGCTTACCAGGGTCTCACAGGCTGATGACAGCTGTGCTGGCATAGGCATGGAGCGATCT
mm4_intron_Mxi1_chr19_53348159 AATCTGGTGTCCCTCTGTGCCAGAGGTACAGCACTGCAGAAGTCATAGGCTAGGACCCTCCTTACGGCAGGGTCCAACCA
TCTGCTGACAGCCTCGCAGG
AGCTGTCAGGCCTCACAGCTCTCTTC
mm4_intergenic_Rnf130|Mir340_chr11_50058302 AGTGGCTTTAAGTCTGGGCTAAAGATGGGTATGGATAGGAACAGAGGTTGACGTAACCAGAGTGGCTGGCCATGGAGGGA
TGGGCGAGGTAATTACAGGC
mm4_intergenic_Mrpl36|Lpcat1_chr13_73416792 AAAGAGGTGGATGTCCGCTGCTGACCCAAATGCAAGTCCCACTACTGACAGCCTCTCTGGGACTTTCCACCCAGGGGCTG
CTGTCCTGAGCTCCCAGAATCCCTAGCTAATGGCAGGTGAGGTG
mm4_exon_Stk11ip_chr1_75529128 TCCAGCCATAGAGGACGACACTAAGGAATCGCCAGAGAAGGTGTCAGAGGAAGGCAGGGTGGAGCCAGAGCC
mm4_intergenic_Wdr20a|Stk30_chr12_110797404 CACTGGACAGTTGGACCCTCCTGTCTGTCCTGCTTGCCCACCGCAGACAACCTTTCTGGTGACTCGAGAAAGCAGCTGCC
CTGTAG
mm4_intron_Rps6kc1_chr1_190723236 GGGAGGGAAGAGCCATGAAAACAGGAAACACAGAGAAGAGAATATNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNCAAG
GAGGCCTGGCCCTGCCGCCAGAGAGGAGGTCAGAGGAAGGGGTCATGAGCTCATCCACCC
mm4_intron_Ppp1r16a_chr15_76671926 GTGAGGGGCCTGCAGAAGGGGCGCCAGCGAGGCGGCCAGCGGAGGGTGGNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNGAAACCTATCAGTGAAGGAAGTTCTTGCAACCTGTTGGTCGGGCAAATCCAT
mm3_intergenic_Spata2|Rnf114/Spata2_chr2_167492119 GGGCCTTCCAAAGCTCTGAAAACAACCTTGGCTGTGCTGGAGCCTGGCTTCCCTCCCCGGAGAGCCTCTCTCGGTTCTGA
AGGGAGCCAAGTCCCCTCTCTA
mm4_intergenic_Mtx3|Cmya5_chr13_92987491 TGTGTGGCTCTTTCCTGTCCCCTCATCTCTTCATAATGCTTCCTAAAAGAGCTGAACTCCGGCACCCCCTTCCCTGGAAA
ACCTCTCTGG
ATCTTGGACTGACGCCGGCATTCCCGGCATCCCCTCCGTGATGCTTA
mm4_intron_Phf17_chr3_41584370 CAGAAGGTGGAGTGAGTTGGACCATAGAGGCTGGCCGAGCAAGGCTGCAAGAGCTGAGGCAGATGGAGGCTCCTTCCCAG
GGTACACACCGGGTTGTCAT
mm4_intergenic_Ilkap|Hes6_chr1_91406008 AGCAAACCTGATCCCTCTGCAACTCAGCAGAACGCGGCCGCTCCTCCTTCGGCTGCCAGCCCCTCTGGCGCCCNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNNTGCTCACGCACAGGTTTCTGGGAG
mm4_exon_Rusc1_chr3_89091214 GCTTCTTACCCACGGCGTGCGCGCGCGGTTCCTCCGGCTCCTGCGAGGCGGATGCCTCCTCACCAGCCCGGGGGCTGGCC
GCCCTGCCCTCCTCCGACAGCCCCTCTGCAATAGGCTGCAGCCCAAGTCGATTTT
mm4_exon_Krt17_chr11_100257640 CGGTTCTCTGTCTCTGCCAGGCTGCCCTCCAGAGATGCTTTCTGCGGGAGGACAGGTGGTAGTCAGTGAGAGTGGCAAAG
CTGACTCC
mm4_exon_Rnf150_chr8_82863914 CAGGGAGTGAGTCGCAGCGACAGGAGAACCAGAGTGGCTGCCNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
NNNNNNNNNNNNNNNNNNNNNNNNNNNACCCCGGAGGAGAGAGATGACA
mm4_intergenic_Parva|2310014F06Rik_chr7_112597411 CTGCCATTTCAGGGTCGGAATATCCAGAGAGGCCCTCTGTGGAGGGGGCTTATATCCCCCATGAGACCCTTTTCTTGCAG
GTCAGGTTCCCATTCGTTTCTCAGGCCAGACC
mm4_intron_Flt1_chr5_147725260 CCTCCTTTGCTCTGGTGGAGCGAGCTCATCGCTAACGACCCCACCTCTGTCCAGCCCCGGGCAGCCTCTCTGACATCTAG
CTTGCTCCAACTAGCCCTTGAGG
mm4_intron_Rapgef1_chr2_29706221 AGCTAATGTGCGATGTGGGTCAGGCCCACAGAACTCCCAAGTAGAGCCCTGCTATGGGTAGTTCCCGGTGCTCAGAACAG
AGCCTGCTGCAGGCAGCCTCTCTGGAACGCAGAGAATCAGCCTTCC
mm4_intron_Masp1_chr16_23496292 CCAGCACCTGGTCTTCTGTTTCTACCTTTGGGGACAAAGTGAAAAGGAATAAAATGAGTGCCTTCCCCTGTCAGCCTCTC
TCG
GGTTCTTCAAGAAGACTGAGAACAGGTCTGCCCACAAGCAGATGAGGCAGCAT
mm4_intergenic_Gm26844|Gm4812_chr13_69316067 AGTGGCTGAGCTGTGATGACCATGTTTCCATCCCCTGACACCTTCTCTGGACATCAGAGACTGTGGTCTATGAGTAATCT
CCTGTAGGCACAAGTTGT
mm4_exon_Ccdc62_chr5_123954507 CAACGTCTACCTGGGCCTTTCCAGCTGCTCAAAACAGCCAGACAGGCTGGACGGGGATGGCCACGACCGCACAGGAACAT
CTGAG
mm4_exon_1700025G04Rik_chr1_151885856 AAGGGCTGCCTGGAATTCTCAAGCAGAGAGGCGCAGCCCTGTGGCTGGAAAAGCCGCCCTCAGCCTCCAGCCTCTCTGGG
GACCTCACCCCATGTGAATCTCTGCC
mm4_intron_Vps8_chr16_21430895 CAAGTCCCTTTGTGCTTGGCAGCCACCACAGAAAGCACCAGTTCCCACTCTTAACAGAACAGAGAGGCTGTTCTGGGAAG
G
CAATGCGAACAACACAGATGTTCACACTGTTCCAC
mm4_intron_Tgfbr3_chr5_107120114 GAGGCAGGAGGTGACACAAGATGAGTCAGTCTGACAGAGAGGCAGTCCTGGGAAGGAACACAGAACGAGAAACCAGGAAG
AGCCTGGTGGAAGTTACAATGGCAGGTCAACCCTTCAGGCCACGAAGCTTA
mm4_intron_Sh3pxd2a_chr19_47342761 CCCGCACCCAGAAAGAATCTTCTAACCCTCTAGGGACAGCCTCTCTGTCAGAGGATAACCTCACTCTTGCTCTTGGGG
mm4_intron_Rwdd2a_chr9_86572362 CTATGCACCTGCGCGAGATGCTGCACCCCACTGCGGTCCGGGGAGGCTGTCCTCCGAGGGGGCCTGCGGATGGGGGCTGG
CGGCGGGGATGACCTAGAGCCCTCCCCTCCGCACTGCAGGCATCCTTCCTAGC
mm4_intron_Mfsd12_chr10_81360837 CCCTGGTGTACCCTAAAGCCAGCTTCCTATGACCTGATTATGCTGGGCCCTCCAGGGACAGCCTCTCTTCAAAGGAACTC
CAGCAGACCAAGTTGTCATGTGATCCCTCTGTTCTGTAGTGGGTGTGTAGCCTGTCTC
mm4_intron_Klhl22_chr16_17783996 TAGAAACACCTGTGGGCAGCCTGTGGCAGACATTTGGGTAGATCCAAACCCTAGAGAGGCTGTACCTGGAAGGGAGATTC
CTTACTTCATTTCCAGGCCACCTTGTGAG
mm4_intron_Cntn1_chr15_92088876 GCCATTTAACCCTGCATGCCCACCCATCCCAGCACAGCCTCTCTGCTCTTCATTGCTGTTATGTTCTTTGTGTTGTTTCT
TCACCACCAATGA
mm4_intron_Arhgap26_chr18_38691553 TAGAGGCCTGATCCGTTCCAAGATCCCCTCCGAGGACTGCCTTTCTGAAGGCAAAGGCCAAGTCTGTAGGAACCA
mm4_intron_Ankrd35_chr3_96689661 GCGGTCTCGGGTGTTCATTAAGCAAATCCAGAGAGGCTGTGTGTGAATGGGAGGAGGAGCCGCTTCTGCTTCTCTTTCTG
GCTCCC
mm4_intron_Akap2_chr4_57808029 CCCCTCTTGACAGGTAGGGATCCCCAGAGAGGCTGTGAGCAAACGGCCTGCCAAGCTGCGCTGTACCTTGAGCCTGGGTC
TCTGACACTTGGGGTG
mm4_intergenic_Pla2g2e|Otud3_chr4_138893379 GCCACCTTGTGAGCTACCATGACATTCTCGGGTAGCAGCCGGGACACAGCCAGGAGGCTGACAGGTTCCCTTCCCAGGAC
AGCTTCTCTGA
GACTGGAGGAACAGTGCCAA
mm4_intergenic_Pard3|mmu-mir-21c_chr8_128054805 GCACACAAATGCAGGGTCAGGACCAGCCAGGAAGAGACTGTACCTTACATTTGGGAAGTTCCCAGAGAGGCTGACCTCTC
AAGG
GTCTTACTCAGGAGACACCCTAAGTGTTCAGTTTGTGGTTGATGTTCCGTCTCA
mm4_intergenic_Hmga2|Gm24298_chr10_120477282 GCCAGCAGTGGGGTTAAAAAGACAGGGCCTAGGCTCTCCTGCCGGATCGCCGCTGGAGGCCACCCCTGAGCGGCTGTCCT
AGGAAGG
CGAAAGAAAATGACTATCTCAGAGTCACCCTGGAGAGCTT
mm4_intergenic_Gm24737|Lhfpl2_chr13_93984681 CATCGAAGGATGGCCCAAGGGCACACCAGAAAGGCTTTCCCAGGATGGCTGGGGCATGCTCATTAGAAGTCTGTTTAATG
AGGGCTGGCACAGTCAGCGAGGGGCAAATGTGAACTCAGC
mm4_intergenic_Gm23120|5S_rRNA_chr11_107851510 GACTTCCCAAGCCTCTGACCCATTTAATCCATCCTCGCACGGCTTCTCTGGCCTAGAATGTCCCTCGCTTTGGTCCAGAC
TT
mm4_intergenic_Gm22670|Gm12434_chr4_47953890 TCTGCCCTGATGCTCTACCTGAGCACAACAGTTTCTCTACAAGAGAAGCTGTCCTCAGAAGGGAAGGAGAGTTAAGTGTG
AGTTTCACAGTAAGTGAAGCATGAAGACAAACACAATTTAACAAGCCAATGCCCTCAAT
mm4_intergenic_Gm14491|Gm14512_chrX_11956264 CTTTCTGTGGCACATGCAGGTGTCCAAATAGGCTGGCCCCGGAAGGCTGGTCCTGGGTCTAGAACTGACACCATGCCTTA
GAAAAGGAAAGCTATCCCAAGGTCAAGACCTCATCAAAGGCCTTCCTGGAGGA
mm4_intergenic_Akt2-ps|Gm12663_chr11_16618265 TAGACACTGGTGGGCTCAGATGCTTCCCAGAGAGGCTGTAACAGGATGGGAGCACCCTAGTGGCCCACCCATCCATTCTT
ATCAACATAGAAAGATGAGATAAGGTACAAGGTCCACTGAGAGGGATCTGTGC
mm4_intergenic_2410088K16Rik|Sh3bp4_chr1_88879641 TGCAGATACAAGCGATGGCATGCCCTGCTTACGGCCCTCCTGGGACTGCCACTCTGGTAAAAATAAAATGCCAATGTTTG
TTGAGGCCTCAGCA
mm4_exon_Lrrc24_chr15_76718359 CTGGACCAAGCCCACAAATGCGCCGCCACGCAATCCCCGAAGCCGGTTGCCAGTGAGCGCCAGTTCAAGCAGGCGAGGCT
GTGCGCGGAAGG
CACCGGACTCCAGCGCACGCAGGGTATTGTTGTG
mm4_exon_Lbh_chr17_72918359 GGAGAGCTTGGTCTGCACTGCCTACGCCGTTGGCAGCGAGGCTGTCTCCGGAAGGCGGACCGGCGCTAGCCCAGTACCCG
CTCTCGGCTGCCCGCTGCTCTCCTCTGCGCCCCTGCCAATCTCATTCA
mm3_intron_Drosha_chr15_12853326 ATTGGTCTGGTGCTCCTGTGCTTCTTCCAGAGACGCTGTCCTCTGATGGCACCGTGCTGATGACGAAGTCA
mm3_intergenic_Fam25c|Sncg_chr14_34357763 CAAAAGTCAGCCCACACAGCCCAAGGTGCTGTTCCTTCCCCCTGCCCTTCAGGCCCTGCTGCCCAGGAACACTGCCCTTC
CTAGGACAGCGTCTCTGG
GTGAGCACTCCAGTGATGTCACTCGACTACCCTTTGCCCAG

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.