Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI9477
Trapped Gene
Zfp71-rs1 (ENSMUSG00000071281)
Vector Insertion
Chr 13: 67811348 - 67811817
Public Clones RRD303 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 11% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000540445 (Chr13:67811818..67811944 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGTGGCCATTGATTTCTCTG Chr13:67811905..67811924 59.65 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000540445 (Chr13:67811818..67811944 -)
Downstram Exon
ENSMUSE00000641321 (Chr13:67811252..67811347 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGTGGCCATTGATTTCTCTG Chr13:67811905..67811924 59.65 45 TGGAGCTCCTCCTCTTTTCA Chr13:67811243..67811262 60.06 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000681337 Chr13:67829998..67830019 No primer for this exon
upstream ENSMUSE00000540445 Chr13:67811818..67811944 TGTGGCCATTGATTTCTCTG Chr13:67811905..67811924 59.65 45

*** Putative Vector Insertion (Chr 13: 67811348 - 67811817) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000641321 Chr13:67811252..67811347 TGGAGCTCCTCCTCTTTTCA Chr13:67811243..67811262 60.06 50
downstream ENSMUSE00000681336 Chr13:67806246..67809866 CCCTAAATGCGGAGAACAAA Chr13:67806419..67806438 60.07 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TACTAATCGCCTTGCAGCAC Chr13:67811749..67811769 59.09 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTACCGTGACTGGGAAAACC Chr13:67811750..67811770 59.31 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000071281