Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI16033
Trapped Gene
Gsta1 (ENSMUSG00000074183)
Vector Insertion
Chr 9: 78080134 - 78082890
Public Clones not available
Private Clones OST467502 (lexicon) OST18010 (lexicon)
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000635393 (Chr9:78080025..78080133 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGCCCGTGCTTCACTACTTC Chr9:78080057..78080076 59.5 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000635393 (Chr9:78080025..78080133 +)
Downstram Exon
ENSMUSE00000635392 (Chr9:78082891..78082942 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGCCCGTGCTTCACTACTTC Chr9:78080057..78080076 59.5 55 TCCAAATCTTCCGGACTCTG Chr9:78082931..78082950 60.19 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000635394 Chr9:78078481..78078537 GCTGGACTGTGAGCTGAGTG Chr9:78078493..78078512 59.76 60
upstream ENSMUSE00000635393 Chr9:78080025..78080133 AGCCCGTGCTTCACTACTTC Chr9:78080057..78080076 59.5 55

*** Putative Vector Insertion (Chr 9: 78080134 - 78082890) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000635392 Chr9:78082891..78082942 TCCAAATCTTCCGGACTCTG Chr9:78082931..78082950 60.19 50
downstream ENSMUSE00000635391 Chr9:78083737..78083869 TGGTGGCGATGTAGTTGAGA Chr9:78083832..78083851 60.26 50
downstream ENSMUSE00000635390 Chr9:78087292..78087433 CAAGGCAGTCTTGGCTTCTC Chr9:78087391..78087410 60.13 55
downstream ENSMUSE00000635389 Chr9:78089334..78089465 CCTGTTGCCCACAAGGTAGT Chr9:78089378..78089397 60.03 55
downstream ENSMUSE00000635388 Chr9:78090260..78090489 GAGGCTGCTGATTCTGCTCT Chr9:78090289..78090308 59.86 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGGTAGGTTGTGCAGGGTTC Chr9:78080133..78080153 59.06 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGGTAGGTTGTGCAGGGTTC Chr9:78080133..78080153 59.06 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000074183