Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25274
Trapped Gene
Pou5f1 (ENSMUSG00000024406)
Vector Insertion
Chr 17: 35646169 - 35646352
Public Clones CMHD-GT_507D7-3 (cmhd)
Private Clones OST189630 (lexicon)
Severity of mutation (?) Insertion after 48% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000142019 (Chr17:35646048..35646168 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCAGAAGAGGATCACCTTGG Chr17:35646104..35646123 59.8 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000142019 (Chr17:35646048..35646168 +)
Downstram Exon
ENSMUSE00000142025 (Chr17:35646353..35646483 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCAGAAGAGGATCACCTTGG Chr17:35646104..35646123 59.8 55 CTCATTGTTGTCGGCTTCCT Chr17:35646474..35646493 60.26 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000142023 Chr17:35643007..35643428 AGCACGAGTGGAAAGCAACT Chr17:35643329..35643348 60.06 50
upstream ENSMUSE00000142019 Chr17:35646048..35646168 GCAGAAGAGGATCACCTTGG Chr17:35646104..35646123 59.8 55

*** Putative Vector Insertion (Chr 17: 35646169 - 35646352) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000142025 Chr17:35646353..35646483 CTCATTGTTGTCGGCTTCCT Chr17:35646474..35646493 60.26 50
downstream ENSMUSE00000142021 Chr17:35646926..35647084 TGATTGGCGATGTGAGTGAT Chr17:35647068..35647087 60.08 45
downstream ENSMUSE00000394011 Chr17:35647233..35647714 TGGGAAAGGTGTCCCTGTAG Chr17:35647337..35647356 59.96 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCCCAGCATGTTCTGATCTC Chr17:35646179..35646199 60.62 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCCCAGCATGTTCTGATCTC Chr17:35646179..35646199 60.62 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024406