Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI12459
Trapped Gene
1810029B16Rik (ENSMUSG00000025591)
Vector Insertion
Chr 8: 69005447 - 69006404
Public Clones F016B01 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 23% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000151985 (Chr8:69006405..69006442 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCCTTGCGTCTCAACCTTAT Chr8:69006407..69006426 59.34 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000151985 (Chr8:69006405..69006442 -)
Downstram Exon
ENSMUSE00000151991 (Chr8:69005362..69005446 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCCTTGCGTCTCAACCTTAT Chr8:69006407..69006426 59.34 50 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000151988 Chr8:69010311..69010406 GAAGAAGCGGACTTGAGAGC Chr8:69010382..69010401 59.31 55
upstream ENSMUSE00000151987 Chr8:69007957..69008069 No primer for this exon
upstream ENSMUSE00000151985 Chr8:69006405..69006442 GCCTTGCGTCTCAACCTTAT Chr8:69006407..69006426 59.34 50

*** Putative Vector Insertion (Chr 8: 69005447 - 69006404) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000151991 Chr8:69005362..69005446 No primer for this exon
downstream ENSMUSE00000151989 Chr8:69004239..69004387 ACTGCTGAACCGATCCAAGT Chr8:69004344..69004363 59.73 50
downstream ENSMUSE00000151986 Chr8:69001914..69001956 No primer for this exon
downstream ENSMUSE00000636652 Chr8:69000245..69000808 GCGCTTCTTAGGAACTGCAT Chr8:69000705..69000724 59.62 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GCCTTGCGTCTCAACCTTAT Chr8:69006405..69006425 59.34 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCCTTGCGTCTCAACCTTAT Chr8:69006405..69006425 59.34 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000025591