Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI34522
Trapped Gene
Polh (ENSMUSG00000023953)
Vector Insertion
Chr 17: 46319633 - 46321759
Public Clones IST12891D8 (tigm) IST12337D6 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 43% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000136620 (Chr17:46321639..46321758 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGGAAAGCTAGGGGCTTCTG Chr17:46321727..46321746 59.98 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000136620 (Chr17:46321639..46321758 -)
Downstram Exon
ENSMUSE00000136619 (Chr17:46319634..46319757 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGGAAAGCTAGGGGCTTCTG Chr17:46321727..46321746 59.98 55 ACCTCGACACATGGCATACA Chr17:46319708..46319727 59.99 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000515197 Chr17:46339346..46339544 GGGAGCCTAGGGCTACCTAC Chr17:46339474..46339493 59.22 65
upstream ENSMUSE00000136615 Chr17:46336341..46336483 No primer for this exon
upstream ENSMUSE00000136618 Chr17:46335583..46335717 CTGGCACAAGTTCGTGAGTC Chr17:46335607..46335626 59.47 55
upstream ENSMUSE00000136621 Chr17:46331151..46331365 CCGGGAAGCTAGTGTTGAAG Chr17:46331343..46331362 59.87 55
upstream ENSMUSE00000136614 Chr17:46327556..46327725 ACTGTGGGAGCCATGATTGT Chr17:46327626..46327645 60.4 50
upstream ENSMUSE00000136616 Chr17:46324976..46325079 CTCTTCAGCCAGATGCCAAT Chr17:46324985..46325004 60.36 50
upstream ENSMUSE00000136620 Chr17:46321639..46321758 AGGAAAGCTAGGGGCTTCTG Chr17:46321727..46321746 59.98 55
upstream ENSMUSE00000136619 Chr17:46319634..46319757 CTTGGCTGTATGCCATGTGT Chr17:46319736..46319755 59.6 50

*** Putative Vector Insertion (Chr 17: 46319633 - 46321759) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000342635 Chr17:46318415..46318480 ACTGCAAAAGCCACCACTGT Chr17:46318437..46318456 60.75 50
downstream ENSMUSE00000498467 Chr17:46312100..46312269 GCTCATCTTGTGAGCGTCAT Chr17:46312134..46312153 58.99 50
downstream ENSMUSE00000410035 Chr17:46308305..46310045 AACTTGGTAGCGCAGAGGAA Chr17:46309982..46310001 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCTAGGGGCTTCTGTCATTG Chr17:46321719..46321739 59.84 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCTAGGGGCTTCTGTCATTG Chr17:46321719..46321739 59.84 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023953