Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI2323
Trapped Gene
Qk (ENSMUSG00000062078)
Vector Insertion
Chr 17: 10466929 - 10475703
Public Clones AE0220 (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 29% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000269846 (Chr17:10475704..10475846 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGGGACCCATTGTTCAGTT Chr17:10475744..10475763 59.68 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000269846 (Chr17:10475704..10475846 -)
Downstram Exon
ENSMUSE00000269819 (Chr17:10466812..10466928 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGGGACCCATTGTTCAGTT Chr17:10475744..10475763 59.68 50 CCAAGGATTCTCCCAACAAA Chr17:10466881..10466900 59.9 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000658322 Chr17:10512187..10512315 CCTGAACTGAGGCGAACTCC Chr17:10512220..10512239 62.28 60
upstream ENSMUSE00000569062 Chr17:10511597..10512245 GCAGCTGATGAACGACAAGA Chr17:10511663..10511682 60.14 50
upstream ENSMUSE00000658321 Chr17:10511597..10511758 GCAGCTGATGAACGACAAGA Chr17:10511663..10511682 60.14 50
upstream ENSMUSE00000658325 Chr17:10511597..10511738 GCAGCTGATGAACGACAAGA Chr17:10511663..10511682 60.14 50
upstream ENSMUSE00000269846 Chr17:10475704..10475846 GTGGGACCCATTGTTCAGTT Chr17:10475744..10475763 59.68 50

*** Putative Vector Insertion (Chr 17: 10466929 - 10475703) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000269819 Chr17:10466812..10466928 CCAAGGATTCTCCCAACAAA Chr17:10466881..10466900 59.9 45
downstream ENSMUSE00000269810 Chr17:10431758..10431901 CTTCAACCGCTCTCTTCAGC Chr17:10431762..10431781 60.28 55
downstream ENSMUSE00000434071 Chr17:10410581..10410668 TGTTGGCGTCTCTGTAGGTG Chr17:10410568..10410587 59.9 55
downstream ENSMUSE00000434057 Chr17:10409025..10409324 TGTAGCTGGTGCCAATGTGT Chr17:10409043..10409062 60.18 50
downstream ENSMUSE00000658324 Chr17:10406224..10406298 GTCTGCGGTCACAATCCTTT Chr17:10406206..10406225 60.12 50
downstream ENSMUSE00000703889 Chr17:10405459..10405472 No primer for this exon
downstream ENSMUSE00000508229 Chr17:10403055..10408339 ACTTTTTGGACACCCAGCAC Chr17:10406146..10406165 60.01 50
downstream ENSMUSE00000351504 Chr17:10403044..10407059 ACTTTTTGGACACCCAGCAC Chr17:10406146..10406165 60.01 50
downstream ENSMUSE00000658323 Chr17:10401473..10402496 GCATGATGCATTGTCACACA Chr17:10402008..10402027 60.13 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTAAGGCCATTCCCTGTTT Chr17:10472719..10472739 59.43 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGTTTTGTTCATGGTTTTCTGC Chr17:10472665..10472687 60.02 36.36 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000062078