Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25352
Trapped Gene
Pigf (ENSMUSG00000024145)
Vector Insertion
Chr 17: 87421291 - 87423004
Public Clones not available
Private Clones OST186981 (lexicon)
Severity of mutation (?) Insertion after 34% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000138675 (Chr17:87423005..87423253 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TTCATTCCGTCCTTCTTCGT Chr17:87423141..87423160 59.67 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000138675 (Chr17:87423005..87423253 -)
Downstram Exon
ENSMUSE00000138683 (Chr17:87421199..87421290 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TTCATTCCGTCCTTCTTCGT Chr17:87423141..87423160 59.67 45 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000138681 Chr17:87424638..87424730 No primer for this exon
upstream ENSMUSE00000138675 Chr17:87423005..87423253 TTCATTCCGTCCTTCTTCGT Chr17:87423141..87423160 59.67 45

*** Putative Vector Insertion (Chr 17: 87421291 - 87423004) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000138683 Chr17:87421199..87421290 No primer for this exon
downstream ENSMUSE00000138685 Chr17:87419741..87419857 AAAACGTTTCCAGCACCAAC Chr17:87419816..87419835 60.01 45
downstream ENSMUSE00000138677 Chr17:87408143..87408251 GCCACGCTCCTGTGAAACTA Chr17:87408165..87408184 61.38 55
downstream ENSMUSE00000138679 Chr17:87396599..87396907 TCCAGAGTGGCGAGATAACC Chr17:87396813..87396832 60.22 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ACTGGCTTTAGGGTGGAGGT Chr17:87422960..87422980 59.99 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACTGGCTTTAGGGTGGAGGT Chr17:87422960..87422980 59.99 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024145