Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI29932
Trapped Gene
Tsc22d1 (ENSMUSG00000022010)
Vector Insertion
Chr 14: 76888483 - 76903635
Public Clones E001C05 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000685772 (Chr14:76888243..76888482 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGTCAGGTTCGAGCATCTGT Chr14:76888293..76888312 60.27 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000685772 (Chr14:76888243..76888482 +)
Downstram Exon
ENSMUSE00000685770 (Chr14:76903636..76903653 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGTCAGGTTCGAGCATCTGT Chr14:76888293..76888312 60.27 55 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000685776 Chr14:76814768..76815524 No primer for this exon
upstream ENSMUSE00000555055 Chr14:76815499..76815524 No primer for this exon
upstream ENSMUSE00000403113 Chr14:76815628..76818816 CTGTGGGAGTGGTTTCCAGT Chr14:76817055..76817074 60 55
upstream ENSMUSE00000555054 Chr14:76815724..76816796 ACTAGCGTTACTCCGGCTCA Chr14:76816229..76816248 60.04 55
upstream ENSMUSE00000685775 Chr14:76815724..76816796 ACTAGCGTTACTCCGGCTCA Chr14:76816229..76816248 60.04 55
upstream ENSMUSE00000706438 Chr14:76815953..76818816 CTGTGGGAGTGGTTTCCAGT Chr14:76817055..76817074 60 55
upstream ENSMUSE00000555053 Chr14:76817043..76818816 CTGTGGGAGTGGTTTCCAGT Chr14:76817055..76817074 60 55
upstream ENSMUSE00000685772 Chr14:76888243..76888482 GGTCAGGTTCGAGCATCTGT Chr14:76888293..76888312 60.27 55

*** Putative Vector Insertion (Chr 14: 76888483 - 76903635) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000685770 Chr14:76903636..76903653 No primer for this exon
downstream ENSMUSE00000122901 Chr14:76904321..76904648 ATCCATCGCCACTGGTCTAC Chr14:76904562..76904581 59.96 55
downstream ENSMUSE00000122900 Chr14:76905056..76905104 GCTACCACACTTGCACCAGA Chr14:76905079..76905098 59.9 55
downstream ENSMUSE00000647340 Chr14:76905056..76905104 GCTACCACACTTGCACCAGA Chr14:76905079..76905098 59.9 55
downstream ENSMUSE00000685769 Chr14:76906224..76907569 TCGCAAGACTTCAGCAGCTA Chr14:76907377..76907396 60.03 50
downstream ENSMUSE00000685771 Chr14:76906224..76906977 TACTGGGGAGAAAGCGAGAA Chr14:76906643..76906662 59.95 50
downstream ENSMUSE00000685774 Chr14:76906224..76907569 TCGCAAGACTTCAGCAGCTA Chr14:76907377..76907396 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGGGTTTGGGTTTGGTTTAG Chr14:76894447..76894467 59.7 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GATTCACTCCCTCCGTGACT Chr14:76888521..76888541 59.1 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

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