Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24141
Trapped Gene
Rpp14 (ENSMUSG00000023156)
Vector Insertion
Chr 14: 8921312 - 8922421
Public Clones (sanger) IST14466E12 (tigm)
Private Clones OST233086 (lexicon) OST214469 (lexicon)
Severity of mutation (?) Insertion after 85% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000133736 (Chr14:8921233..8921311 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAAGCAAGAAGTGTGCCTTCA Chr14:8921280..8921300 60.04 42.86 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000133736 (Chr14:8921233..8921311 +)
Downstram Exon
ENSMUSE00000394747 (Chr14:8922422..8924350 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAAGCAAGAAGTGTGCCTTCA Chr14:8921280..8921300 60.04 42.86 AGCTTCCGGGACCATAACTT Chr14:8923059..8923078 59.96 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000306272 Chr14:8912881..8913026 CAGTGATCCTCTCCGTCCTG Chr14:8912978..8912997 60.82 60
upstream ENSMUSE00000306266 Chr14:8916222..8916308 CAAGAGCCCCTCAGAGTATCA Chr14:8916270..8916290 59.44 52.38
upstream ENSMUSE00000306256 Chr14:8916431..8916515 CAAACAGCTGCTCGTCTCAG Chr14:8916473..8916492 59.92 55
upstream ENSMUSE00000133739 Chr14:8919978..8920054 AGAAGACACTGTCGGCCATC Chr14:8920018..8920037 60.27 55
upstream ENSMUSE00000133736 Chr14:8921233..8921311 AAAGCAAGAAGTGTGCCTTCA Chr14:8921280..8921300 60.04 42.86

*** Putative Vector Insertion (Chr 14: 8921312 - 8922421) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000394747 Chr14:8922422..8924350 AGCTTCCGGGACCATAACTT Chr14:8923059..8923078 59.96 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGCCTTCAGAGTGATTCAGG Chr14:8921294..8921314 58.96 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGCCTTCAGAGTGATTCAGG Chr14:8921294..8921314 58.96 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023156