Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI22169
Trapped Gene
AC182749.2-203 (ENSMUSG00000079709)
Vector Insertion
Chr 17: 15118642 - 15119263
Public Clones IST11661G7 (tigm) IST12266G10 (tigm)
Private Clones OST305142 (lexicon) OST185774 (lexicon)
Severity of mutation (?) Insertion after 80% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000703661 (Chr17:15118540..15118641 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTCGGCGTGATGAAACTAGC Chr17:15118594..15118613 60.93 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000703661 (Chr17:15118540..15118641 +)
Downstram Exon
ENSMUSE00000703660 (Chr17:15119264..15119353 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTCGGCGTGATGAAACTAGC Chr17:15118594..15118613 60.93 55 GCATGTCCAAAGACTTGTGC Chr17:15119352..15119371 59.3 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000716076 Chr17:15115829..15115863 No primer for this exon
upstream ENSMUSE00000658235 Chr17:15116465..15116633 TTACCAGGGGATTCCATGAC Chr17:15116468..15116487 59.6 50
upstream ENSMUSE00000703662 Chr17:15117222..15117361 GGCGAAGTCTGGACTACAGG Chr17:15117222..15117241 59.87 60
upstream ENSMUSE00000703661 Chr17:15118540..15118641 CTCGGCGTGATGAAACTAGC Chr17:15118594..15118613 60.93 55

*** Putative Vector Insertion (Chr 17: 15118642 - 15119263) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000703660 Chr17:15119264..15119353 GCATGTCCAAAGACTTGTGC Chr17:15119352..15119371 59.3 50
downstream ENSMUSE00000658227 Chr17:15119853..15119873 No primer for this exon
downstream ENSMUSE00000703658 Chr17:15121782..15121879 TGCCACAAGACGTTTCTCAG Chr17:15121849..15121868 60.03 50
downstream ENSMUSE00000703657 Chr17:15122738..15122874 GGTGACCTTTGTGTGGTGAA Chr17:15122819..15122838 59.42 50
downstream ENSMUSE00000703656 Chr17:15123682..15123796 TTCCCAATATGGCAACATGA Chr17:15123770..15123789 59.74 40
downstream ENSMUSE00000703652 Chr17:15124560..15124662 AACATGGTGTAGTCGGCAAA Chr17:15124583..15124602 59.05 45
downstream ENSMUSE00000703649 Chr17:15125038..15125067 No primer for this exon
downstream ENSMUSE00000703648 Chr17:15125862..15125930 CAAGGAAGCTGGTTCATGCT Chr17:15125911..15125930 60.4 50
downstream ENSMUSE00000658240 Chr17:15127511..15127768 GTTGATCTCCCCATGGCTTA Chr17:15127588..15127607 59.89 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGCGACGTGAGTGTGAGAAT Chr17:15118637..15118657 61.29 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000079709