Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI15131
Trapped Gene
Rps2 (ENSMUSG00000044533)
Vector Insertion
Chr 17: 24858660 - 24858747
Public Clones PST12397-NR (escells) PSTVUpb14c1 (vanderbilt)
Private Clones not available
Severity of mutation (?) Insertion after 91% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000465206 (Chr17:24858568..24858659 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TACCTGACTCCCGACCTCTG Chr17:24858606..24858625 60.25 60 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000465206 (Chr17:24858568..24858659 +)
Downstram Exon
ENSMUSE00000497807 (Chr17:24858748..24858868 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TACCTGACTCCCGACCTCTG Chr17:24858606..24858625 60.25 60 TCTGGTGTGGGTTTTCACAA Chr17:24858786..24858805 59.98 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000628947 Chr17:24857055..24857397 ACTAGCAAATGGCGGATGAC Chr17:24857213..24857232 60.1 50
upstream ENSMUSE00000138837 Chr17:24857479..24857676 CCTGCCCATTAAGGAGTCTG Chr17:24857556..24857575 59.69 55
upstream ENSMUSE00000478496 Chr17:24857914..24858247 TCGTCGCTATTGGGGACTAC Chr17:24857918..24857937 60.1 55
upstream ENSMUSE00000465206 Chr17:24858568..24858659 TACCTGACTCCCGACCTCTG Chr17:24858606..24858625 60.25 60

*** Putative Vector Insertion (Chr 17: 24858660 - 24858747) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000497807 Chr17:24858748..24858868 TCTGGTGTGGGTTTTCACAA Chr17:24858786..24858805 59.98 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAGGTATTCGGATTCGGTTG Chr17:24858658..24858678 60.32 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAGGTATTCGGATTCGGTTG Chr17:24858658..24858678 60.32 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000044533