Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI21654
Trapped Gene
Cacng6 (ENSMUSG00000078815)
Vector Insertion
Chr 7: 3425754 - 3431155
Public Clones IST13920F4 (tigm)
Private Clones OST322893 (lexicon) OST179299 (lexicon)
Severity of mutation (?) Insertion after 42% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000677464 (Chr7:3425423..3425753 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAGAATCTCGGCTTGACTCC Chr7:3425501..3425520 59.95 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000677464 (Chr7:3425423..3425753 +)
Downstram Exon
ENSMUSE00000677463 (Chr7:3431156..3431230 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAGAATCTCGGCTTGACTCC Chr7:3425501..3425520 59.95 55 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000677464 Chr7:3425423..3425753 CAGAATCTCGGCTTGACTCC Chr7:3425501..3425520 59.95 55

*** Putative Vector Insertion (Chr 7: 3425754 - 3431155) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000677463 Chr7:3431156..3431230 No primer for this exon
downstream ENSMUSE00000677462 Chr7:3432537..3432674 GCAGGGACTCTGCACCTTTA Chr7:3432644..3432663 60.4 55
downstream ENSMUSE00000677461 Chr7:3435420..3435658 CAAGGTGAGCAGGAGGAAAC Chr7:3435607..3435626 59.84 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGAGGTGAGAGGCTGTGTTC Chr7:3428751..3428771 59.84 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGAGGTGAGAGGCTGTGTTC Chr7:3428751..3428771 59.84 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000078815