Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25877
Trapped Gene
Gm525 (ENSMUSG00000072553)
Vector Insertion
Chr 11: 88950006 - 88954199
Public Clones CMHD-GT_455E5-3 (cmhd) PSTVUpb17f11 (vanderbilt)
Private Clones OST155191 (lexicon) OST37565 (lexicon) OST26246 (lexicon) OST8569 (lexicon)
OST5764 (lexicon) OST1791 (lexicon)
Severity of mutation (?) Insertion after 100% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000648988 (Chr11:88949870..88950005 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CGCACTGTGACAGGAACATT Chr11:88949952..88949971 59.75 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000648988 (Chr11:88949870..88950005 +)
Downstram Exon
ENSMUSE00000648986 (Chr11:88954200..88954354 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CGCACTGTGACAGGAACATT Chr11:88949952..88949971 59.75 50 CTGGCGTTCTGCCTACACTT Chr11:88954256..88954275 60.45 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000648991 Chr11:88935238..88935319 GGCATCATGGAGAAGCTGTT Chr11:88935259..88935278 60.23 50
upstream ENSMUSE00000648990 Chr11:88936303..88936403 GCAAAACAGCTCCTGAGGTC Chr11:88936332..88936351 60 55
upstream ENSMUSE00000648988 Chr11:88949870..88950005 CGCACTGTGACAGGAACATT Chr11:88949952..88949971 59.75 50

*** Putative Vector Insertion (Chr 11: 88950006 - 88954199) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000648986 Chr11:88954200..88954354 CTGGCGTTCTGCCTACACTT Chr11:88954256..88954275 60.45 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAGGGATAATCGCCTTGCAG Chr11:88953051..88953071 60.19 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTAGGGACGTGACTGGGAAA Chr11:88950050..88950070 59.97 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000072553