Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI29847
Trapped Gene
Pmm1 (ENSMUSG00000022474)
Vector Insertion
Chr 15: 81788325 - 81791295
Public Clones (sanger) D170F02 (ggtc) E047A02 (ggtc) (ggtc) D170F02 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 11% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000394648 (Chr15:81791145..81791294 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CATCCTCTGCCTGTTTGACG Chr15:81791174..81791193 61.8 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000394648 (Chr15:81791145..81791294 -)
Downstram Exon
ENSMUSE00000680688 (Chr15:81788326..81788340 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CATCCTCTGCCTGTTTGACG Chr15:81791174..81791193 61.8 55 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000394648 Chr15:81791145..81791294 CATCCTCTGCCTGTTTGACG Chr15:81791174..81791193 61.8 55
upstream ENSMUSE00000680688 Chr15:81788326..81788340 No primer for this exon

*** Putative Vector Insertion (Chr 15: 81788325 - 81791295) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000127320 Chr15:81788223..81788340 CTGCTCGGCGATCTTAGAGT Chr15:81788220..81788239 59.74 55
downstream ENSMUSE00000127321 Chr15:81786792..81786868 GCCGTCCGTGTTTATACTGC Chr15:81786784..81786803 60.53 55
downstream ENSMUSE00000127330 Chr15:81786599..81786690 GGCCATGTAGCTAAGGCAGA Chr15:81786600..81786619 60.37 55
downstream ENSMUSE00000398536 Chr15:81786079..81786178 CAGTTCCGAGAACTCGATCC Chr15:81786063..81786082 59.8 55
downstream ENSMUSE00000127325 Chr15:81783155..81783230 TTCTCCCGGATCTTCTCCTT Chr15:81783189..81783208 60.15 50
downstream ENSMUSE00000127332 Chr15:81782354..81782469 TCCAGGCAGTAGCGCTTATC Chr15:81782390..81782409 60.51 55
downstream ENSMUSE00000484589 Chr15:81781542..81782042 GGGTCCGCATAGATCTCAAA Chr15:81781989..81782008 60.04 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr15:81788303..81788323 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGAGGCGTGACTGGGAAAA Chr15:81788309..81788329 63.61 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022474