Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI26296
Trapped Gene
Rwdd2b (ENSMUSG00000041079)
Vector Insertion
Chr 16: 87437377 - 87437538
Public Clones CMHD-GT_469C10-3 (cmhd)
Private Clones OST126398 (lexicon)
Severity of mutation (?) Insertion after 24% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000266745 (Chr16:87437539..87437759 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAATGTGAGCCTGGACCTCT Chr16:87437553..87437572 60.26 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000266745 (Chr16:87437539..87437759 -)
Downstram Exon
ENSMUSE00000266735 (Chr16:87437309..87437376 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAATGTGAGCCTGGACCTCT Chr16:87437553..87437572 60.26 55 CTGTAATTTCGGGCAGGACT Chr16:87437291..87437310 59.19 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000471614 Chr16:87440682..87440837 TCTCTCTGGGGTGAGACCTG Chr16:87440737..87440756 60.39 60
upstream ENSMUSE00000266745 Chr16:87437539..87437759 CAATGTGAGCCTGGACCTCT Chr16:87437553..87437572 60.26 55

*** Putative Vector Insertion (Chr 16: 87437377 - 87437538) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000266735 Chr16:87437309..87437376 CTGTAATTTCGGGCAGGACT Chr16:87437291..87437310 59.19 50
downstream ENSMUSE00000266722 Chr16:87436818..87437180 GTGAAAGCGAGATCCTCTGG Chr16:87436958..87436977 59.95 55
downstream ENSMUSE00000443798 Chr16:87433576..87434871 CAGTGAGCCAACAGGTCTGA Chr16:87434579..87434598 60.02 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAATGTGAGCCTGGACCTCT Chr16:87437551..87437571 60.26 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAATGTGAGCCTGGACCTCT Chr16:87437551..87437571 60.26 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000041079