Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI21548
Trapped Gene
Pfdn5 (ENSMUSG00000001289)
Vector Insertion
Chr 15: 102157281 - 102158952
Public Clones not available
Private Clones OST326301 (lexicon) OST302667 (lexicon) OST54190 (lexicon)
Severity of mutation (?) Insertion after 45% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000133101 (Chr15:102157249..102157280 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000133101 (Chr15:102157249..102157280 +)
Downstram Exon
ENSMUSE00000133104 (Chr15:102158953..102159027 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000368771 Chr15:102156610..102156691 No primer for this exon
upstream ENSMUSE00000133103 Chr15:102156864..102156966 No primer for this exon
upstream ENSMUSE00000133101 Chr15:102157249..102157280 No primer for this exon

*** Putative Vector Insertion (Chr 15: 102157281 - 102158952) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000133104 Chr15:102158953..102159027 No primer for this exon
downstream ENSMUSE00000133099 Chr15:102159143..102159248 No primer for this exon
downstream ENSMUSE00000283367 Chr15:102161704..102161918 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGGTCCCACTGACGAGTTCT Chr15:102157262..102157282 60.71 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCACGGGAAATGCTACCTTA Chr15:102157291..102157311 59.95 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000001289