Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI21586
Trapped Gene
Pmm1 (ENSMUSG00000022474)
Vector Insertion
Chr 15: 81786179 - 81786598
Public Clones Ayu21-T149 (egtc)
Private Clones OST325070 (lexicon)
Severity of mutation (?) Insertion after 32% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000127330 (Chr15:81786599..81786690 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TTAGCTACATGGCCCTGCTC Chr15:81786616..81786635 60.37 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000127330 (Chr15:81786599..81786690 -)
Downstram Exon
ENSMUSE00000398536 (Chr15:81786079..81786178 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TTAGCTACATGGCCCTGCTC Chr15:81786616..81786635 60.37 55 CAGTTCCGAGAACTCGATCC Chr15:81786063..81786082 59.8 55

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
upstream ENSMUSE00000127320 Chr15:81788223..81788340 ACTCTAAGATCGCCGAGCAG Chr15:81788242..81788261 59.74 55
upstream ENSMUSE00000127321 Chr15:81786792..81786868 GGACGGCTACTCTCCAAACA Chr15:81786793..81786812 60.26 55
upstream ENSMUSE00000127330 Chr15:81786599..81786690 TTAGCTACATGGCCCTGCTC Chr15:81786616..81786635 60.37 55

*** Putative Vector Insertion (Chr 15: 81786179 - 81786598) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
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 TACATGGCCCTGCTCAGACT Chr15:81786609..81786629 60.82 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TACATGGCCCTGCTCAGACT Chr15:81786609..81786629 60.82 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022474