Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI35567
Trapped Gene
5430437P03Rik (ENSMUSG00000031820)
Vector Insertion
Chr 8: 73920920 - 73921577
Public Clones (ggtc) IST14153D9 (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
ENSMUSE00000366419 (Chr8:73920760..73920919 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGTCGCCGTGGTCTAACTT Chr8:73920876..73920895 60.18 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000366419 (Chr8:73920760..73920919 +)
Downstram Exon
ENSMUSE00000396414 (Chr8:73921578..73921924 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGTCGCCGTGGTCTAACTT Chr8:73920876..73920895 60.18 55 ACCCTTGGTGTTCGAATCTG Chr8:73921908..73921927 59.97 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000366419 Chr8:73920760..73920919 GTGTCGCCGTGGTCTAACTT Chr8:73920876..73920895 60.18 55

*** Putative Vector Insertion (Chr 8: 73920920 - 73921577) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000396414 Chr8:73921578..73921924 ACCCTTGGTGTTCGAATCTG Chr8:73921908..73921927 59.97 50
downstream ENSMUSE00000213343 Chr8:73922291..73922349 CCTCCGAAAGATCCAGACAG Chr8:73922318..73922337 59.8 55
downstream ENSMUSE00000221895 Chr8:73923425..73923545 CAAATTCGTGGCTCTTGTCA Chr8:73923516..73923535 59.84 45
downstream ENSMUSE00000392428 Chr8:73923628..73923706 CCGTCTCCAGGTCATACAGG Chr8:73923691..73923710 60.52 60
downstream ENSMUSE00000682507 Chr8:73925579..73925603 GCTGAAGAGGCCTTCCAAAT Chr8:73925601..73925620 60.71 50
downstream ENSMUSE00000221875 Chr8:73926645..73926774 GGGATGGTTTGCACATTCTC Chr8:73926695..73926714 60.33 50
downstream ENSMUSE00000213344 Chr8:73926875..73926961 CCTCAGTGCCATTGTGGATA Chr8:73926938..73926957 59.52 50
downstream ENSMUSE00000379280 Chr8:73928094..73928448 AATAGTGTGCCTGCGCTCTC Chr8:73928356..73928375 60.57 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTGTCGCCGTGGTCTAACTT Chr8:73920877..73920897 60.18 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGTCGCCGTGGTCTAACTT Chr8:73920877..73920897 60.18 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031820