Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33364
Trapped Gene
Praf2 (ENSMUSG00000031149)
Vector Insertion
Chr X: 7305889 - 7306841
Public Clones IST14227H5 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 33% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000206889 (ChrX:7305697..7305888 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GATCCACAACGATGGTGTCA ChrX:7305794..7305813 60.39 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000206889 (ChrX:7305697..7305888 +)
Downstram Exon
ENSMUSE00000206888 (ChrX:7306842..7307059 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GATCCACAACGATGGTGTCA ChrX:7305794..7305813 60.39 50 CCACTACCAGTGCACTCAGG ChrX:7306888..7306907 59.33 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000206889 ChrX:7305697..7305888 GATCCACAACGATGGTGTCA ChrX:7305794..7305813 60.39 50

*** Putative Vector Insertion (Chr X: 7305889 - 7306841) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000206888 ChrX:7306842..7307059 CCACTACCAGTGCACTCAGG ChrX:7306888..7306907 59.33 60
downstream ENSMUSE00000242962 ChrX:7307394..7308184 GGCTAGGAAAAGGGTTGGTC ChrX:7307606..7307625 59.94 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GAGGTAGCGATTGGCTGAAG ChrX:7305900..7305920 59.98 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAGGTAGCGATTGGCTGAAG ChrX:7305900..7305920 59.98 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031149