Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI33148
Trapped Gene
Nme2 (ENSMUSG00000020857)
Vector Insertion
Chr 11: 93816808 - 93817196
Public Clones IST14531C11 (tigm) IST14531D11 (tigm) IST12362E5 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000351001 (Chr11:93817140..93817195 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000351001 (Chr11:93817140..93817195 -)
Downstram Exon
ENSMUSE00000492655 (Chr11:93816809..93817097 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000351001 Chr11:93817140..93817195 No primer for this exon
upstream ENSMUSE00000110580 Chr11:93816809..93816938 No primer for this exon
upstream ENSMUSE00000492655 Chr11:93816809..93817097 No primer for this exon

*** Putative Vector Insertion (Chr 11: 93816808 - 93817196) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000473358 Chr11:93814114..93814215 No primer for this exon
downstream ENSMUSE00000460702 Chr11:93813214..93813326 No primer for this exon
downstream ENSMUSE00000412222 Chr11:93811131..93811360 No primer for this exon
downstream ENSMUSE00000110587 Chr11:93811130..93811360 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTCTTTGCTTCTGTCGCTCA Chr11:93817169..93817189 59.47 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTCTTTGCTTCTGTCGCTCA Chr11:93817169..93817189 59.47 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000020857