Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI35053
Trapped Gene
D19Wsu162e (ENSMUSG00000047731)
Vector Insertion
Chr 19: 46673784 - 46681201
Public Clones (ggtc) IST12411A10 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000708213 (Chr19:46673627..46673783 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGGAAGAAGAGAGGGAGGAG Chr19:46673630..46673649 59.37 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000708213 (Chr19:46673627..46673783 +)
Downstram Exon
ENSMUSE00000641158 (Chr19:46681202..46681249 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGGAAGAAGAGAGGGAGGAG Chr19:46673630..46673649 59.37 60 CGAACCAGTCCCACTGTCTT Chr19:46681242..46681261 60.15 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000708213 Chr19:46673627..46673783 GGGAAGAAGAGAGGGAGGAG Chr19:46673630..46673649 59.37 60
upstream ENSMUSE00000713184 Chr19:46673627..46673783 GGGAAGAAGAGAGGGAGGAG Chr19:46673630..46673649 59.37 60

*** Putative Vector Insertion (Chr 19: 46673784 - 46681201) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000641158 Chr19:46681202..46681249 CGAACCAGTCCCACTGTCTT Chr19:46681242..46681261 60.15 55
downstream ENSMUSE00000689283 Chr19:46698043..46698069 TCTAAGCCCCCAGAGGAAAG Chr19:46698069..46698088 60.69 55
downstream ENSMUSE00000471790 Chr19:46718843..46718945 GCAGCACTGAGACTGTCCAC Chr19:46718926..46718945 59.61 60
downstream ENSMUSE00000706656 Chr19:46718843..46719057 CCCCTGGACAAGGACTTACA Chr19:46718967..46718986 59.96 55
downstream ENSMUSE00000472696 Chr19:46726964..46727125 AAACGGCAACGCTGAGTAGT Chr19:46727121..46727140 59.94 50
downstream ENSMUSE00000389933 Chr19:46728455..46731875 CTGCAAATCTGACCGTAGCA Chr19:46731686..46731705 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGGTAAGGGGAGGGCTTCTG Chr19:46679783..46679803 61.84 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCCTGTCAGTGAGGGCTGAG Chr19:46673756..46673776 63.37 65 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000047731