Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32595
Trapped Gene
EG433481 (ENSMUSG00000074758)
Vector Insertion
Chr 2: 144000262 - 144004885
Public Clones IST11408C12 (tigm) IST12387H12 (tigm) IST12838G3 (tigm) IST10713F3 (tigm)
IST10066G12 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 16% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000640508 (Chr2:144004764..144004884 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGTTCTGCCGAGACCACAGT Chr2:144004829..144004848 59.91 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000640508 (Chr2:144004764..144004884 -)
Downstram Exon
ENSMUSE00000640506 (Chr2:144000263..144000480 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGTTCTGCCGAGACCACAGT Chr2:144004829..144004848 59.91 55 TGAGAAATGGTGCGTCTCTG Chr2:144000356..144000375 59.98 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000682462 Chr2:144014877..144015026 AGATCTGCATGCCATTTTGA Chr2:144014882..144014901 59.23 40
upstream ENSMUSE00000640508 Chr2:144004764..144004884 AGTTCTGCCGAGACCACAGT Chr2:144004829..144004848 59.91 55
upstream ENSMUSE00000640506 Chr2:144000263..144000480 CAGAGACGCACCATTTCTCA Chr2:144000378..144000397 59.98 50

*** Putative Vector Insertion (Chr 2: 144000262 - 144004885) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000640505 Chr2:143998886..144000139 AGGGAGTTTCGACAGCTCAA Chr2:143999297..143999316 59.99 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCATTTTAATCGCCTTGCAG Chr2:144001820..144001840 60.58 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCATTTCGTGACTGGGAAAA Chr2:144001820..144001840 60.86 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000074758