Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI37238
Trapped Gene
Dctd (ENSMUSG00000031562)
Vector Insertion
Chr 8: 49222729 - 49223532
Public Clones CMHD-GT_472D11-3 (cmhd)
Private Clones not available
Severity of mutation (?) Insertion after 67% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000210765 (Chr8:49222612..49222728 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCCGTGTAACGAATGTGCTA Chr8:49222691..49222710 59.61 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000210765 (Chr8:49222612..49222728 +)
Downstram Exon
ENSMUSE00000636947 (Chr8:49223533..49223606 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCCGTGTAACGAATGTGCTA Chr8:49222691..49222710 59.61 50 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000359191 Chr8:49195539..49195602 No primer for this exon
upstream ENSMUSE00000399702 Chr8:49197013..49197144 CAAAGAAGCAAGGACCCAAG Chr8:49197118..49197137 59.85 50
upstream ENSMUSE00000210767 Chr8:49197322..49197457 TTGTTGGGATCGGGTACAAT Chr8:49197356..49197375 60.05 45
upstream ENSMUSE00000210765 Chr8:49222612..49222728 CCCGTGTAACGAATGTGCTA Chr8:49222691..49222710 59.61 50

*** Putative Vector Insertion (Chr 8: 49222729 - 49223532) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000636947 Chr8:49223533..49223606 No primer for this exon
downstream ENSMUSE00000210773 Chr8:49224136..49224232 CTCCTCGCTGTCGTGGTATT Chr8:49224188..49224207 60.28 55
downstream ENSMUSE00000371021 Chr8:49225697..49227017 GACGGGTGAGAAGATTCCAA Chr8:49225968..49225987 60.05 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GATTTGTCACGTCTGCATCG Chr8:49222751..49222771 60.27 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GATTTGTCACGTCTGCATCG Chr8:49222751..49222771 60.27 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031562