Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI803
Trapped Gene
Trim71 (ENSMUSG00000079259)
Vector Insertion
Chr 9: 114424986 - 114434071
Public Clones (sanger) AA0094 (sanger) (sanger) AN0208 (sanger) (sanger)
CH0260 (sanger) (sanger) RRS310 (baygenomics) (ggtc) 5SP143E12 (ggtc)
(ggtc) (ggtc) 5SH004C06 (ggtc) (ggtc) (ggtc) IST12376B6 (tigm)
IST12376B6 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 38% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000689095 (Chr9:114434072..114434239 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGTGACACCTGCTCTGTCC Chr9:114434206..114434225 59.44 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000689095 (Chr9:114434072..114434239 -)
Downstram Exon
ENSMUSE00000689094 (Chr9:114424851..114424985 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGTGACACCTGCTCTGTCC Chr9:114434206..114434225 59.44 60 TCCACAACAGCTCACACTCC Chr9:114424831..114424850 59.87 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000689096 Chr9:114471423..114473487 GGGGAGTCGTGCAAAATAAA Chr9:114473347..114473366 59.94 45
upstream ENSMUSE00000689095 Chr9:114434072..114434239 CTGTGACACCTGCTCTGTCC Chr9:114434206..114434225 59.44 60

*** Putative Vector Insertion (Chr 9: 114424986 - 114434071) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000689094 Chr9:114424851..114424985 TCCACAACAGCTCACACTCC Chr9:114424831..114424850 59.87 55
downstream ENSMUSE00000689092 Chr9:114420393..114423214 ATTCTGCTGGTCGCTCACTT Chr9:114422713..114422732 60.02 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGTGGGTAGTAATCGCCTTG Chr9:114434009..114434029 59.45 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGAAGTTGCTGTGTGTGAGG Chr9:114434034..114434054 59.31 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CCCAGTATTCCACACAAACG Chr9:114434209..114434229 58.9 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CCATCTGCAGAGAGTCGTGA Chr9:114434184..114434204 60.14 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000079259