Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6407
Trapped Gene
B3gnt2 (ENSMUSG00000051650)
Vector Insertion
Chr 11: 22737196 - 22759493
Public Clones YHC081 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000370857 (Chr11:22759494..22759703 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGCTGGAGGTGTCCCTAGA Chr11:22759498..22759517 59.4 60 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000370857 (Chr11:22759494..22759703 -)
Downstram Exon
ENSMUSE00000357467 (Chr11:22734865..22737195 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGCTGGAGGTGTCCCTAGA Chr11:22759498..22759517 59.4 60 GCCCAGCAACTTGACTCTTC Chr11:22737129..22737148 60 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000331247 Chr11:22760218..22760336 AACCTGAAGGTCCGAAGAGG Chr11:22760292..22760311 60.62 55
upstream ENSMUSE00000370857 Chr11:22759494..22759703 GAGCTGGAGGTGTCCCTAGA Chr11:22759498..22759517 59.4 60

*** Putative Vector Insertion (Chr 11: 22737196 - 22759493) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000357467 Chr11:22734865..22737195 GCCCAGCAACTTGACTCTTC Chr11:22737129..22737148 60 55
downstream ENSMUSE00000493466 Chr11:22734857..22737195 GCCCAGCAACTTGACTCTTC Chr11:22737129..22737148 60 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCTCTTGTGCCTACCACTCC Chr11:22759456..22759476 59.87 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTCCTGTGTGCTCTGGTACG Chr11:22759440..22759460 59.49 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000051650