Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI9167
Trapped Gene
Wdr6 (ENSMUSG00000066357)
Vector Insertion
Chr 9: 108478914 - 108480824
Public Clones RRE091 (baygenomics) IST10544D8 (tigm) IST13682G11 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 3% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000331581 (Chr9:108480825..108480957 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TCATACTCCTCCCGGTTACG Chr9:108480859..108480878 59.95 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000331581 (Chr9:108480825..108480957 -)
Downstram Exon
ENSMUSE00000240681 (Chr9:108476432..108478913 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TCATACTCCTCCCGGTTACG Chr9:108480859..108480878 59.95 55 AAAGCAGCTCCTCGTAACCA Chr9:108477635..108477654 60.01 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000331581 Chr9:108480825..108480957 TCATACTCCTCCCGGTTACG Chr9:108480859..108480878 59.95 55

*** Putative Vector Insertion (Chr 9: 108478914 - 108480824) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000240681 Chr9:108476432..108478913 AAAGCAGCTCCTCGTAACCA Chr9:108477635..108477654 60.01 50
downstream ENSMUSE00000240659 Chr9:108476236..108476331 CCAGGCCTATCGTTGTCAAG Chr9:108476265..108476284 60.65 55
downstream ENSMUSE00000240629 Chr9:108476025..108476141 GCTGGTTAGGTGCCTCATGT Chr9:108476007..108476026 60.14 55
downstream ENSMUSE00000221376 Chr9:108475820..108475938 GGCTAGGCTGCCATCTGTAG Chr9:108475874..108475893 60 60
downstream ENSMUSE00000514800 Chr9:108474649..108475744 GCTGGTCTATGGAGGCTGAG Chr9:108475426..108475445 59.97 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGGCTTGGTCGGCTCTAGTC Chr9:108480800..108480820 60.92 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTCTCGTGACTGGGAAAACC Chr9:108480757..108480777 59.7 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 GAGAACGCAGCTCTCTTCTTG Chr9:108480955..108480976 59.51 52.38 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 TGTGACTCTGGCTGGTTCCT Chr9:108477934..108477954 60.86 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000066357