Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25299
Trapped Gene
Gm525 (ENSMUSG00000072553)
Vector Insertion
Chr 11: 88936404 - 88949869
Public Clones IST12140C11 (tigm) IST12140C11 (tigm)
Private Clones OST188600 (lexicon) OST111189 (lexicon)
Severity of mutation (?) Insertion after 58% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000648990 (Chr11:88936303..88936403 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCAAAACAGCTCCTGAGGTC Chr11:88936332..88936351 60 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000648990 (Chr11:88936303..88936403 +)
Downstram Exon
ENSMUSE00000648988 (Chr11:88949870..88950005 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCAAAACAGCTCCTGAGGTC Chr11:88936332..88936351 60 55 CCAGGTCCTTCACACAGGAT Chr11:88949998..88950017 59.96 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

*** Putative Vector Insertion (Chr 11: 88936404 - 88949869) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000648988 Chr11:88949870..88950005 CCAGGTCCTTCACACAGGAT Chr11:88949998..88950017 59.96 55
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 AAATGGTAATCGCCTTGCAG Chr11:88936449..88936469 60.1 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACTACCGTGACTGGGAAAACC Chr11:88936450..88936471 60.27 52.38 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000072553