Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI17180
Trapped Gene
Ndufb3 (ENSMUSG00000026032)
Vector Insertion
Chr 1: 58643515 - 58647933
Public Clones not available
Private Clones OST448074 (lexicon) OST365289 (lexicon) OST352885 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000337266 (Chr1:58643443..58643514 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTCTCCGGCTGTAAGGTTGA Chr1:58643460..58643479 60.26 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000337266 (Chr1:58643443..58643514 +)
Downstram Exon
ENSMUSE00000155114 (Chr1:58647934..58648093 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTCTCCGGCTGTAAGGTTGA Chr1:58643460..58643479 60.26 55 AGCTTCTTCTGCACCGTTTC Chr1:58648063..58648082 59.62 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000337266 Chr1:58643443..58643514 GTCTCCGGCTGTAAGGTTGA Chr1:58643460..58643479 60.26 55

*** Putative Vector Insertion (Chr 1: 58643515 - 58647933) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000155114 Chr1:58647934..58648093 AGCTTCTTCTGCACCGTTTC Chr1:58648063..58648082 59.62 50
downstream ENSMUSE00000343428 Chr1:58652493..58652792 ATCACCATTCTGGGAATCCA Chr1:58652637..58652656 60.13 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAGGGAGCTGGTGGGAGACT Chr1:58643522..58643542 61.18 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TAGGGAGCTGGTGGGAGACT Chr1:58643522..58643542 61.18 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000026032