Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33185
Trapped Gene
2010100O12Rik (ENSMUSG00000067847)
Vector Insertion
Chr 2: 155970326 - 155971337
Public Clones IST13570D5 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 55% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000680878 (Chr2:155969969..155970325 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ATTCGGAGTGAGACGTCGAG Chr2:155969972..155969991 60.41 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000680878 (Chr2:155969969..155970325 +)
Downstram Exon
ENSMUSE00000553460 (Chr2:155971338..155971519 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ATTCGGAGTGAGACGTCGAG Chr2:155969972..155969991 60.41 55 TTAGGTATTGCAGGGCATCC Chr2:155971478..155971497 59.92 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000680878 Chr2:155969969..155970325 ATTCGGAGTGAGACGTCGAG Chr2:155969972..155969991 60.41 55
upstream ENSMUSE00000680877 Chr2:155969973..155970021 GAGTGAGACGTCGAGCTGAG Chr2:155969977..155969996 58.86 60
upstream ENSMUSE00000712669 Chr2:155969976..155970016 GAGTGAGACGTCGAGCTGAG Chr2:155969977..155969996 58.86 60
upstream ENSMUSE00000680881 Chr2:155969980..155970016 No primer for this exon
upstream ENSMUSE00000719558 Chr2:155970181..155970325 GGCACCTTCTCCTGTCTCAG Chr2:155970306..155970325 59.99 60
upstream ENSMUSE00000553461 Chr2:155970195..155970325 GGCACCTTCTCCTGTCTCAG Chr2:155970306..155970325 59.99 60
upstream ENSMUSE00000712760 Chr2:155970195..155970325 GGCACCTTCTCCTGTCTCAG Chr2:155970306..155970325 59.99 60

*** Putative Vector Insertion (Chr 2: 155970326 - 155971337) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000553460 Chr2:155971338..155971519 TTAGGTATTGCAGGGCATCC Chr2:155971478..155971497 59.92 50
downstream ENSMUSE00000680879 Chr2:155971338..155971529 TTAGGTATTGCAGGGCATCC Chr2:155971478..155971497 59.92 50
downstream ENSMUSE00000706115 Chr2:155971338..155971530 TTAGGTATTGCAGGGCATCC Chr2:155971478..155971497 59.92 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTAATCGCCTTGCAGCACAT Chr2:155970376..155970396 61.32 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCACCTTCTCCTGTCTCAGGT Chr2:155970308..155970329 60.84 57.14 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000067847