Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI31377
Trapped Gene
Adarb2 (ENSMUSG00000052551)
Vector Insertion
Chr 13: 8671766 - 8696865
Public Clones IST10259D4 (tigm) IST14245G3 (tigm) IST12061F4 (tigm) IST14123F10 (tigm)
IST13013C9 (tigm) IST11842G4 (tigm) IST13172H7 (tigm) IST10952D7 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 54% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000476554 (Chr13:8671651..8671765 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CGGCCTGACATCTGTGTATG Chr13:8671707..8671726 60.14 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000476554 (Chr13:8671651..8671765 +)
Downstram Exon
ENSMUSE00000441669 (Chr13:8696866..8697034 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CGGCCTGACATCTGTGTATG Chr13:8671707..8671726 60.14 55 CCACGATCTCAGCATGACAG Chr13:8696985..8697004 60.43 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000471563 Chr13:8202467..8202566 AGAGGGTCTGGAGGGCTAAG Chr13:8202491..8202510 59.83 60
upstream ENSMUSE00000468343 Chr13:8558350..8558436 TGTCAACCTTCCTTGCTCCT Chr13:8558365..8558384 59.84 50
upstream ENSMUSE00000465454 Chr13:8568913..8569820 AAACCGAAATGTGAGCAACC Chr13:8568938..8568957 59.98 45
upstream ENSMUSE00000476554 Chr13:8671651..8671765 CGGCCTGACATCTGTGTATG Chr13:8671707..8671726 60.14 55

*** Putative Vector Insertion (Chr 13: 8671766 - 8696865) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000441669 Chr13:8696866..8697034 CCACGATCTCAGCATGACAG Chr13:8696985..8697004 60.43 55
downstream ENSMUSE00000441655 Chr13:8700826..8700977 TTTCTCGCAGCCTGTAACCT Chr13:8700899..8700918 60.01 50
downstream ENSMUSE00000441646 Chr13:8712846..8713014 CTGACGATGTGTTTGCTGCT Chr13:8712873..8712892 60.06 50
downstream ENSMUSE00000441662 Chr13:8731036..8731217 AGGCCTGTTTTGTCGGTATG Chr13:8731210..8731229 59.99 50
downstream ENSMUSE00000455934 Chr13:8751819..8751997 TGAAGTGGGGAGACTTTCCT Chr13:8751869..8751888 58.72 50
downstream ENSMUSE00000684878 Chr13:8756472..8759554 GCATAGGCCAAAGTGATGGT Chr13:8758877..8758896 59.96 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCCCCCATTTTATAGGTCA Chr13:8689719..8689739 58.39 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AGCCCCCATTTTATAGGTCA Chr13:8689719..8689739 58.39 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000052551