Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI27251
Trapped Gene
Rab1b (ENSMUSG00000024870)
Vector Insertion
Chr 19: 5104929 - 5105177
Public Clones not available
Private Clones OST65729 (lexicon)
Severity of mutation (?) Insertion after 15% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000145703 (Chr19:5105178..5105250 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGCTTTTGATTGGTGACTCC Chr19:5105214..5105234 59.73 47.62 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000145703 (Chr19:5105178..5105250 -)
Downstram Exon
ENSMUSE00000145700 (Chr19:5104833..5104928 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGCTTTTGATTGGTGACTCC Chr19:5105214..5105234 59.73 47.62 ACACCGATGGTGCTGATGTA Chr19:5104866..5104885 59.99 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000145701 Chr19:5106925..5107019 GGAGTGGCAACCATCTTGGA Chr19:5106994..5107013 63.37 55
upstream ENSMUSE00000145703 Chr19:5105178..5105250 CTGCTTTTGATTGGTGACTCC Chr19:5105214..5105234 59.73 47.62

*** Putative Vector Insertion (Chr 19: 5104929 - 5105177) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000145700 Chr19:5104833..5104928 ACACCGATGGTGCTGATGTA Chr19:5104866..5104885 59.99 50
downstream ENSMUSE00000145698 Chr19:5104641..5104736 CCGCGATAGTAGCTGGAAGT Chr19:5104659..5104678 59.5 55
downstream ENSMUSE00000553253 Chr19:5100667..5100798 ACGACCTTCTTGGTGGTGAG Chr19:5100664..5100683 60.15 55
downstream ENSMUSE00000379928 Chr19:5099207..5100572 TGCTGTCGATCTTCAGGTTG Chr19:5100391..5100410 59.98 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTGCGGTTTGCTGTGAGTAA Chr19:5105168..5105188 60.05 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGCGGTTTGCTGTGAGTAA Chr19:5105168..5105188 60.05 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024870