Inturned fh347 K108X ENU male 7149
>wt seq
AAATCTGAAGCAAAACTCAAAAAGCTGACCAAGATCTTGAGACGGAAGCGGCGACCATC
TC
>fh347 seq
AAATCTGAAGCAAAACTCAAAAAGCTGACCTAGATCTTGAGACGGAAGCGGCGACCATC
TC
1 mismatch:
Cutting wild type forward sequence:
ENZYME
RECOGNITION SEQUENCE
PRIMER SEQUENCE
BalI :
TGGCCA
AAATCTGAAGCAAAACTC
BclI :
TGATCA
AAATCTGAAGCAAAACTC
CfrI :
YGGCCR
AAATCTGAAGCAAAACTC
MboII :
GAAGA
AAATCTGAAGCAAAACTC
Cutting wild type reverse sequence:
ENZYME
RECOGNITION SEQUENCE
PRIMER SEQUENCE
BstXI :
CCANNNNNNTGG
GAGATGGTCGCCGCTTCCG
NlaIII :
CATG
GAGATGGTCGCCGCTTCCG
PflMI :
CCANNNNNTGG
GAGATGGTCGCCGCTTCCG
Cutting mutant forward sequence:
ENZYME
RECOGNITION SEQUENCE
PRIMER SEQUENCE
AluI :
AGCT
AAATCTGAAGCAAAACTC
CviJI :
RGCY
AAATCTGAAGCAAAACTC
MaeII :
ACGT
AAATCTGAAGCAAAACTC
XbaI :
TCTAGA
AAATCTGAAGCAAAACTC
Cutting mutant reverse sequence:
ENZYME
RECOGNITION SEQUENCE
PRIMER SEQUENCE
AvrII :
CCTAGG
GAGATGGTCGCCGCTTCCG
TaqI :
TCGA
GAGATGGTCGCCGCTTCCG
> Dr. inturned +/- 300 bp around fh347 K108*
TGAAATGGCCCCTCACCAATTAGAGTTTGAGGCCCCTGCTCTAACGTCTTTTCCTCTGTG
TTGTTTCGCAGTGATCTGGAGCCGGAATGGCTGGATGACGTCCAGAAGAACGGTCAGCTT
TTCTACCTGGAGCTGAGTGATGGAGAAGAAGAGGCTCTACTGGCTCAGGTCTCCGCCAAT
CACTCCGCTGCCACCAATCACGTGCGCTTCAGCGAAAAGGAGGCGGAGATTATCACTGAC
AATGCAAAAAGGCAGGCAAACTCCAGCAACAAATCTGAAGCAAAACTCAAAAAGCTGAC
CAAGATCTTGAGACGGAAGCGGCGACCATCTCAGAGGAAAGCTGAGGGAAAAGACTCTT
CCCAGCGTCCCGCTTCCATCCTGAAGAATCAGGCGGGACAGAGGCCTGGAGTGGTGGTTC
AGCAGCAGCGTCTGAAGGATGTGTGTGTGTACCTGAACCCAAAACGCCTCAGCAGCGTCT
CGTCTTCATCTGCTGACAGAGGCGGGCTGCTGGAGGCTTTACTGGGGGTCGTGCACAGAC
CTGGAGGAAATACAGGGAAAAGAGGAGGAAAACTCATCATCCATGGACTCATCCCTCAC
AGTC
GT282
>inturned_fh347_geno_DCAP_A_f
AAATCTGAAGCAAAACTCAAAAAGCTGATC
GT283
> inturned_fh347_geno_DCAP_A_r
GTTCAGGTACACACACACATCCTTCAG
After PCR, cut with Xba1
+/+:
185 bp bands
fh347/+ :
185,160,25 bp
fh347/fh347: 160,25 bp