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Contents:

PCR primers for off-targets of CCACGCTCCCGGGAAGTCGG 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_52314003_F TCGTCGGCAGCGTCAACCGCCAGAGAAGATGGTG 60.0 1.00
ontarget_mm0_exon_5730457N03Rik/Evx1_chr6_52314003_R GTCTCGTGGGCTCGGCTGCCTGCTAGTCCATCGAC 60.2 1.00
mm3_intron_Inpp4a_chr1_37366459_F TCGTCGGCAGCGTCTGTCATGGGTTCACGAGGTG 59.9 0.69
mm3_intron_Inpp4a_chr1_37366459_R GTCTCGTGGGCTCGGTCAGTAGAAGGACGCTTGGC 59.7 0.69
mm4_intron_Styxl1_chr5_135774770_F Primer3: not found at this Tm N.d. 0.50
mm4_intron_Styxl1_chr5_135774770_R Primer3: not found at this Tm N.d. 0.50
mm4_intergenic_Rpsa-ps10|Mir3963_chr3_150677303_F TCGTCGGCAGCGTCACGTCTAAATGGCAAGCGGT 60.3 0.48
mm4_intergenic_Rpsa-ps10|Mir3963_chr3_150677303_R GTCTCGTGGGCTCGGGTCCTGAAAAGGCATGCGTG 60.1 0.48
mm4_intergenic_Rpl31-ps18|Ins1_chr19_51923168_F TCGTCGGCAGCGTCTGGTTGGGCTTTATGCAGGA 59.5 0.42
mm4_intergenic_Rpl31-ps18|Ins1_chr19_51923168_R GTCTCGTGGGCTCGGTTTTGCAGGGACTCAGAGCA 59.5 0.42
mm4_intergenic_Cxcr3|Tpt1-ps6_chrX_101752753_F Primer3: not found at this Tm N.d. 0.38
mm4_intergenic_Cxcr3|Tpt1-ps6_chrX_101752753_R Primer3: not found at this Tm N.d. 0.38
mm4_intergenic_Gm23342|Myt1l_chr12_29049731_F TCGTCGGCAGCGTCTGCAATATGTCCTTGTTACGACT 58.1 0.38
mm4_intergenic_Gm23342|Myt1l_chr12_29049731_R GTCTCGTGGGCTCGGTCATAAACCAGTCCAGGGCC 59.3 0.38
mm4_intron_Ntmt1_chr2_30820796_F Primer3: not found at this Tm N.d. 0.37
mm4_intron_Ntmt1_chr2_30820796_R Primer3: not found at this Tm N.d. 0.37
mm4_intergenic_Gm22825|Zfhx3_chr8_108099352_F Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Gm22825|Zfhx3_chr8_108099352_R Primer3: not found at this Tm N.d. 0.33
mm4_intergenic_Lipc|Aqp9_chr9_70963548_F TCGTCGGCAGCGTCAATGAGAGCAGCTTCCGCTC 60.4 0.33
mm4_intergenic_Lipc|Aqp9_chr9_70963548_R GTCTCGTGGGCTCGGACTCCAAGACACAGTGCCTG 59.8 0.33
mm4_exon_Mmadhc_chr2_50296417_F TCGTCGGCAGCGTCGCAGTATAGGCGTGTCCCTC 59.9 0.27
mm4_exon_Mmadhc_chr2_50296417_R GTCTCGTGGGCTCGGAGTGCCAGCGCCCTAGTG 62.0 0.27
mm4_intergenic_Tmem30b|Prkch_chr12_73580600_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Tmem30b|Prkch_chr12_73580600_R Primer3: not found at this Tm N.d. 0.22
mm4_exon_Dis3l2_chr1_86703848_F Primer3: not found at this Tm N.d. 0.22
mm4_exon_Dis3l2_chr1_86703848_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Maml3|Foxo1/Gm20402_chr3_52267896_F Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Maml3|Foxo1/Gm20402_chr3_52267896_R Primer3: not found at this Tm N.d. 0.22
mm4_intergenic_Igdcc3|Gm25313_chr9_65188730_F TCGTCGGCAGCGTCAAGTTGGTGCAGGAGAGCAG 60.2 0.21
mm4_intergenic_Igdcc3|Gm25313_chr9_65188730_R GTCTCGTGGGCTCGGCAAGGTGGCAGCAAAAGTCC 59.9 0.21
mm4_intron_Itga2b_chr11_102455343_F TCGTCGGCAGCGTCCAGGAAACTCTGAGGGCAGG 60.0 0.19
mm4_intron_Itga2b_chr11_102455343_R GTCTCGTGGGCTCGGTTGTGCTGGCCATGTACCAT 59.9 0.19
mm4_intergenic_Agpat2|Fam69b_chr2_26609717_F TCGTCGGCAGCGTCGCCCTGTCTGGTAGGCATTT 60.0 0.17
mm4_intergenic_Agpat2|Fam69b_chr2_26609717_R GTCTCGTGGGCTCGGAGCTGTTGAAGGCACCAGTT 60.1 0.17
mm4_intergenic_D730003I15Rik|Nenf_chr1_191268706_F TCGTCGGCAGCGTCGCCTCCTTGTGTACTCCTGC 60.3 0.17
mm4_intergenic_D730003I15Rik|Nenf_chr1_191268706_R GTCTCGTGGGCTCGGGGGCATGATCCCATCAGAAGA 59.5 0.17
mm4_intron_Dis3l2_chr1_86772418_F TCGTCGGCAGCGTCTCTTGAAATGTCTTGCTGTGCA 59.0 0.16
mm4_intron_Dis3l2_chr1_86772418_R GTCTCGTGGGCTCGGAAGTCATGCACCCAACCAGC 61.1 0.16
mm4_intron_Galc_chr12_98234435_F TCGTCGGCAGCGTCCCACACTGTGTAGGTTCCAGG 60.2 0.15
mm4_intron_Galc_chr12_98234435_R GTCTCGTGGGCTCGGAGCAAACCTGGGCGAAGAG 60.3 0.15
mm4_intergenic_Nsmce2|Trib1_chr15_59636198_F TCGTCGGCAGCGTCATCAAAGTGGGTTCAGGGCG 60.6 0.15
mm4_intergenic_Nsmce2|Trib1_chr15_59636198_R GTCTCGTGGGCTCGGCTGTCCCATGGCTTCCTCAG 60.1 0.15
mm4_intergenic_Gm26474|RP24-351J24.2_chr5_120313450_F TCGTCGGCAGCGTCTCCCCAGAGGAAGAGTGGAG 59.9 0.13
mm4_intergenic_Gm26474|RP24-351J24.2_chr5_120313450_R GTCTCGTGGGCTCGGCCATTAGCCTGCTCAGTCCC 60.1 0.13
mm4_intergenic_Gm25509|n-R5-8s1_chr18_73294214_F TCGTCGGCAGCGTCTGTCTCCAAATCCTGCCCAA 59.2 0.13
mm4_intergenic_Gm25509|n-R5-8s1_chr18_73294214_R GTCTCGTGGGCTCGGGAAGATGGGGTTGGCTCAGG 60.3 0.13
mm4_intergenic_Otogl|Ppp1r12a_chr10_107951530_F Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Otogl|Ppp1r12a_chr10_107951530_R Primer3: not found at this Tm N.d. 0.12
mm4_intergenic_Muc6|Muc2_chr7_141694688_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Muc6|Muc2_chr7_141694688_R Primer3: not found at this Tm N.d. 0.11
mm4_exon_Txlna_chr4_129640255_F TCGTCGGCAGCGTCGAAAAGCAGAGCTGGCACAC 60.0 0.11
mm4_exon_Txlna_chr4_129640255_R GTCTCGTGGGCTCGGGACCCAGACAGACAGCTCCT 60.9 0.11
mm4_intergenic_Nfyb|1700028I16Rik_chr10_82777904_F Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Nfyb|1700028I16Rik_chr10_82777904_R Primer3: not found at this Tm N.d. 0.11
mm4_intergenic_Suz12|Crlf3_chr11_80043438_F Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Suz12|Crlf3_chr11_80043438_R Primer3: not found at this Tm N.d. 0.10
mm4_intergenic_Sec11c|Grp_chr18_65852082_F Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Sec11c|Grp_chr18_65852082_R Primer3: not found at this Tm N.d. 0.09
mm4_intergenic_Slc4a1|Bloodlinc_chr11_102373811_F TCGTCGGCAGCGTCGCCTCACCTATCCTCCCTGA 60.1 0.08
mm4_intergenic_Slc4a1|Bloodlinc_chr11_102373811_R GTCTCGTGGGCTCGGCTTCCTAAGGCAGAGGGAGC 59.5 0.08
mm4_intergenic_Arap3|Pcdh1_chr18_38106270_F Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Arap3|Pcdh1_chr18_38106270_R Primer3: not found at this Tm N.d. 0.07
mm4_intergenic_Tfcp2l1/Gm23497|Gli2_chr1_118801853_F TCGTCGGCAGCGTCGCTGCCTTTTACCCTCCCAT 60.0 0.05
mm4_intergenic_Tfcp2l1/Gm23497|Gli2_chr1_118801853_R GTCTCGTGGGCTCGGCTGCTGAAAAGTGCCAACCC 59.9 0.05
mm4_intron_Gpc6_chr14_117579856_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Gpc6_chr14_117579856_R Primer3: not found at this Tm N.d. 0.05
mm4_intron_Dscam_chr16_97039342_F Primer3: not found at this Tm N.d. 0.05
mm4_intron_Dscam_chr16_97039342_R Primer3: not found at this Tm N.d. 0.05
mm4_exon_Nbl1_chr4_139083274_F Primer3: not found at this Tm N.d. 0.04
mm4_exon_Nbl1_chr4_139083274_R Primer3: not found at this Tm N.d. 0.04
mm4_intron_Dnajc13_chr9_104236432_F TCGTCGGCAGCGTCGTAAAGCCCATGCACGGTTG 60.1 0.04
mm4_intron_Dnajc13_chr9_104236432_R GTCTCGTGGGCTCGGGTGCAGTCCTGACTTTCCCA 59.8 0.04
mm4_intergenic_Gm25423|Gm26054_chr7_116746092_F Primer3: not found at this Tm N.d. 0.04
mm4_intergenic_Gm25423|Gm26054_chr7_116746092_R Primer3: not found at this Tm N.d. 0.04
mm4_intron_Adrm1_chr2_180171942_F Primer3: not found at this Tm N.d. 0.03
mm4_intron_Adrm1_chr2_180171942_R Primer3: not found at this Tm N.d. 0.03
mm4_exon_Gpatch4_chr3_88043511_F TCGTCGGCAGCGTCAACGATTCTCAAGGGCTGGG 60.0 0.03
mm4_exon_Gpatch4_chr3_88043511_R GTCTCGTGGGCTCGGGCTGTAAGCCTTCCCCACAT 60.0 0.03
mm4_intron_Vps13b_chr15_35914111_F TCGTCGGCAGCGTCCAGCTCCTGGCAAGTCTGAA 59.9 0.03
mm4_intron_Vps13b_chr15_35914111_R GTCTCGTGGGCTCGGAACATCGGTGACAGGGACAC 59.9 0.03
mm4_intergenic_Gm25781|Ipo5_chr14_120814497_F Primer3: not found at this Tm N.d. 0.03
mm4_intergenic_Gm25781|Ipo5_chr14_120814497_R Primer3: not found at this Tm N.d. 0.03
mm4_intron_Sptbn1_chr11_30161423_F TCGTCGGCAGCGTCGTTGGCAGCGACAATGGAAG 60.1 0.03
mm4_intron_Sptbn1_chr11_30161423_R GTCTCGTGGGCTCGGAGGACTGGATAACGCTGCTG 59.8 0.03
mm4_intergenic_Gm24839|Tmem132c_chr5_126756040_F Primer3: not found at this Tm N.d. 0.02
mm4_intergenic_Gm24839|Tmem132c_chr5_126756040_R Primer3: not found at this Tm N.d. 0.02
mm3_exon_Thumpd3_chr6_113046262_F TCGTCGGCAGCGTCCGTTCTTCCTCTTCCGGGTC 60.1 0.02
mm3_exon_Thumpd3_chr6_113046262_R GTCTCGTGGGCTCGGAAGTCAGGCCTCGGAGGG 60.6 0.02
mm3_intergenic_Tug1/TUG1_2/TUG1_1|Tug1_chr11_3648646_F TCGTCGGCAGCGTCCTCCAACTGGGTCCTAGCAG 59.4 0.02
mm3_intergenic_Tug1/TUG1_2/TUG1_1|Tug1_chr11_3648646_R GTCTCGTGGGCTCGGGGTCGTCCGGGTGGTTGG 62.6 0.02
mm4_intron_Rpap3_chr15_97705535_F TCGTCGGCAGCGTCTCAAGCCAAGCCTGCATACC 60.6 0.02
mm4_intron_Rpap3_chr15_97705535_R GTCTCGTGGGCTCGGCAGGGTTCGGCCTGAGCT 61.7 0.02
mm4_intergenic_1700011I03Rik|Slc12a2_chr18_57878616_F Primer3: not found at this Tm N.d. 0.00
mm4_intergenic_1700011I03Rik|Slc12a2_chr18_57878616_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Ptgs1_chr2_36230593_F TCGTCGGCAGCGTCCATGAGTCGTGAGTCCGACC 60.1 0.00
mm4_exon_Ptgs1_chr2_36230593_R GTCTCGTGGGCTCGGACCAGAGCGAGAGACTCCTT 59.9 0.00
mm4_exon_Gm12827/Rnf220_chr4_117496956_F Primer3: not found at this Tm N.d. 0.00
mm4_exon_Gm12827/Rnf220_chr4_117496956_R Primer3: not found at this Tm N.d. 0.00
mm4_exon_Dlec1_chr9_119121909_F TCGTCGGCAGCGTCTCATGCCCCAAAAGAGCTGG 60.6 0.00
mm4_exon_Dlec1_chr9_119121909_R GTCTCGTGGGCTCGGGTTTCTGGTCCCAGGTGTGG 60.5 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_52314003 AACCGCCAGAGAAGATGGTGCCCCACGGTTGCCTGAGCCCGCGAGCCGGCCCTCCGACTTCCCGGGAGCGTGGCGGGGGA
GGCCAGGAGGAGGAGCCGGTCGATGGACTAGCAGGCAG
mm3_intron_Inpp4a_chr1_37366459 TGTCATGGGTTCACGAGGTGAGGGTGGGTCGGTTATAGGAACCACCGACTTCCCTGGATTGTGGTACGTGGGTCAGAGGG
ACGCCAAGCGTCCTTCTACTGA
mm4_intergenic_Rpsa-ps10|Mir3963_chr3_150677303 ACGTCTAAATGGCAAGCGGTTCTCAAGCTCCAGGGAAGTCTGAGGTGAGATGGGTTATTAAGAGAAGTATAAGGTGCAGA
AGTTAAGGTGGTGGGCACGCATGCCTTTTCAGGAC
mm4_intergenic_Rpl31-ps18|Ins1_chr19_51923168 TGGTTGGGCTTTATGCAGGAGTAATAGCCACGTTCCTGCAAAGTCGGTGGTGATTCGGTTTTTAAATATTGAATTCTACT
TAAAATGAACTATGATGCACAGTAATGCTCTGAGTCCCTGCAAAA
mm4_intergenic_Gm23342|Myt1l_chr12_29049731 TGCAATATGTCCTTGTTACGACTTTCCCATCAACTTCCCGGGAGTGTTGTATGTTATTAGGCTTTCTATCTCAGGGCAAA
AGGCTTTTGTTTGCTGTTTTTTTTTTTTAAAGGCCCTGGACTGGTTTATGA
mm4_intergenic_Lipc|Aqp9_chr9_70963548 AATGAGAGCAGCTTCCGCTCAGGCTGCAGCAGAGGCTTGGCTCCTCCAGCTTCCCAGGAGGGTGGAAAGCTCAAAGCCAG
TTCTGGCTAGTGTGGATTAGAGTTCCCAGCAGCCCAGGCACTGTGTCTTGGAGT
mm4_exon_Mmadhc_chr2_50296417 GCAGTATAGGCGTGTCCCTCCCCAAAGCCCGGCCTCGGGCCCCGCGTCCCCGGGAAGCCGGGGGACACTAGGGCGCTGGC
ACT
mm4_intergenic_Igdcc3|Gm25313_chr9_65188730 AAGTTGGTGCAGGAGAGCAGCAACTGAGGCAGAAGGCTCTCCTCTGACTTCCCTGGAGCCAGGCCTGGCAGTCTGGTAGC
CCTGCACTTAAAAAGCTGGACCGCGACATGAGGACTTTTGCTGCCACCTTG
mm4_intron_Itga2b_chr11_102455343 CAGGAAACTCTGAGGGCAGGTCTCAGTTTCCTCCGACTTCCCGACAGCAGGGGAAGGGATCTGAAGGCCTGAGGATTTCA
GACGTCCTGCTTGGGGGGATGGGAGGGATGGTACATGGCCAGCACAA
mm4_intergenic_Agpat2|Fam69b_chr2_26609717 GCCCTGTCTGGTAGGCATTTAAGCATGTGGCTGTTGTCCTCCAATCCCCTCCAACTTCCCTGCAGCCTGGTCTGGGACAG
ACAGCAGGCTGTTTCCCAACCTCTAGCTATAACTGGTGCCTTCAACAGCT
mm4_intergenic_D730003I15Rik|Nenf_chr1_191268706 GCCTCCTTGTGTACTCCTGCTTCCTCTGTAATTCAAGGAGTTAACTCAGCAGCCCCACTCTTCTGGGAAGTCAGCGGCGG
CCGCTCCTCAGCCCATAGCCTTCTTCTGATGGGATCATGCCC
mm4_intron_Dis3l2_chr1_86772418 TCTTGAAATGTCTTGCTGTGCAAGAAAGTCTAATATTAACATGTAAGCTTTGGAGAGCCATAGACTTCCCTGGAGAGTGG
TAGTCACATACTAAAATATCCCAGCTGGTTGGGTGCATGACTT
mm4_intron_Galc_chr12_98234435 CCACACTGTGTAGGTTCCAGGATAGTGAGCTCTAAGAGAAAGGGTACAAGAGGAGAAAAGTCAGAAGCCCCACACTTCCG
TGAAGTAGGTGG
GTGAACTCAAATACAGCTCTTCGCCCAGGTTTGCT
mm4_intergenic_Nsmce2|Trib1_chr15_59636198 ATCAAAGTGGGTTCAGGGCGCATACCTGTAGCCCTCCGTCTTCCTGGGCCCGTGGAGGGATCTCCTGCCAACCTTGTAGT
CTGAGGAAGCCATGGGACAG
mm4_intergenic_Gm26474|RP24-351J24.2_chr5_120313450 TCCCCAGAGGAAGAGTGGAGGGGTATTCTAGAGTAGTTCCCTCCAAGTCAGCCCCCTGCCATGCTCATGGGCAGTCGGGG
G
AGCAGGCCACCGCCTGAGCGCCCAGAGGGAGGGACTGAGCAGGCTAATGG
mm4_intergenic_Gm25509|n-R5-8s1_chr18_73294214 TGTCTCCAAATCCTGCCCAATCATCAGAATCTGAACACAATTGCAACTGTAGATCATAGGTGATCTTCCCTCAGAATTCA
CCCTCAATACCCCCGACTCCCCCTGAGCTTGGGTAGCATCCTGAGCCAACCCCATCTTC
mm4_exon_Txlna_chr4_129640255 GAAAAGCAGAGCTGGCACACACCCTCGGTCTTCCCAGGAGCCTGGCTTGTGGAACCTTCGGCTTCCGCCCCAGGAGCTGT
CTGTCTGGGTC
mm4_intergenic_Slc4a1|Bloodlinc_chr11_102373811 GCCTCACCTATCCTCCCTGAATGCAGTGGGAATGACACTAAGTATGACAAAGCAGGAACTGCATTCCCCCCTGGCCCATC
CTCCCCACCCCCACTCTCCCTGGAAATCGTGGGGGGGGGGGCTCCCTCTGCCTTAGGAAG
mm4_intergenic_Tfcp2l1/Gm23497|Gli2_chr1_118801853 GCTGCCTTTTACCCTCCCATCTTGGCTCCCTGCTTCTAGGTGGTTAGAGCCTGAGACAGCCAGGCTCTCAGGAAGTGGGA
GG
AAGATGGGGGTTGGCACTTTTCAGCAG
mm4_intron_Dnajc13_chr9_104236432 GTAAAGCCCATGCACGGTTGAGAAATGGGATGCATCATGAGAAAACCCAAAAGTCCTCCATCCCCGCTCCCTGGAAGTCT
CTGG
TTTAATGGAGAGGCCTGGCTGGGAAAGTCAGGACTGCAC
mm4_exon_Gpatch4_chr3_88043511 AACGATTCTCAAGGGCTGGGAAATAAAGCTTGCCTCCGCCTTCCGGGAAGTGTGGTTTTGAATTACTGTGATGAGAAATA
GAATGTGGGGAAGGCTTACAGC
mm4_intron_Vps13b_chr15_35914111 CAGCTCCTGGCAAGTCTGAATTTAAATAGGAAAGCAGACCCAAAGCCACGCTCCTGGGACATGGGTGGGGGAGCACTTCC
CTCTCGCTAACCTAATCAGTGTGTCCCTGTCACCGATGTT
mm4_intron_Sptbn1_chr11_30161423 GTTGGCAGCGACAATGGAAGACAATCCAGAGGGTTTCCAAGCTCAAGCTGTTGCAGCTGGCCAAGCTCAGGGGAAGGCGG
TGG
GAGGACAGGGAGCAGCAGCGTTATCCAGTCCT
mm3_exon_Thumpd3_chr6_113046262 CGTTCTTCCTCTTCCGGGTCAGAGTGCACCCAACTGCGCACAGCTCTCCGCGCACACTCCCCCGCCTACCCCGGAGCGTG
G
GAGCACCGTGCCCTCCGAGGCCTGACTT
mm3_intergenic_Tug1/TUG1_2/TUG1_1|Tug1_chr11_3648646 CTCCAACTGGGTCCTAGCAGGCCCCGAGGGGTGGCGCCAGGCCTCCAGGCCAGCCCCGGCCACCCCAGCTTCCCCCGCCG
CCTCCCCGGGAGCGGGG
CGGAGTTGCCCGCCGCCGCCAACCACCCGGACGACC
mm4_intron_Rpap3_chr15_97705535 TCAAGCCAAGCCTGCATACCCTGGCCTGACGGAGCCCGCAGGTCCGCCGCCCTCGCGGGACCGTGGCTCCGGAGGTCCCG
CCACCGCAGACCGCCCGAGCCCGGACGCGGAGCCAGCTCAGGCCGAACCCTG
mm4_exon_Ptgs1_chr2_36230593 CATGAGTCGTGAGTCCGACCCCAGTGGTGCCCCCACGCGTCCCGGAATTCGGTGGCCTGCAGGCGGAGCCTTGAACGCTA
GGCTCAACTCTCTCTTCCTTCTGCAGGAAGGAGTCTCTCGCTCTGGT
mm4_exon_Dlec1_chr9_119121909 TCATGCCCCAAAAGAGCTGGCCGCCACCGAGTTTCCGGGTGAGTGGCCGTGGACCCCGACCGGGGAAAGGAACGGAACCC
ACCCAGGCAACTGAGCTTGGGGGAGGCCACTGAGCCACACCTGGGACCAGAAAC

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.