Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI27393
Trapped Gene
Polr3g (ENSMUSG00000035834)
Vector Insertion
Chr 13: 81833777 - 81838742
Public Clones not available
Private Clones OST60776 (lexicon)
Severity of mutation (?) Insertion after 17% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000411409 (Chr13:81838743..81838892 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AATGGCTGGCAATAAAGGAA Chr13:81838841..81838860 59.55 40 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000411409 (Chr13:81838743..81838892 -)
Downstram Exon
ENSMUSE00000461631 (Chr13:81833647..81833776 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AATGGCTGGCAATAAAGGAA Chr13:81838841..81838860 59.55 40 GTTTTTCTTCGGGTGGTTCA Chr13:81833628..81833647 59.95 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000411101 Chr13:81849857..81850012 GTAGAATCTGCGGCTCCTTG Chr13:81849928..81849947 59.98 55
upstream ENSMUSE00000411409 Chr13:81838743..81838892 AATGGCTGGCAATAAAGGAA Chr13:81838841..81838860 59.55 40

*** Putative Vector Insertion (Chr 13: 81833777 - 81838742) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000461631 Chr13:81833647..81833776 GTTTTTCTTCGGGTGGTTCA Chr13:81833628..81833647 59.95 45
downstream ENSMUSE00000248747 Chr13:81828196..81828255 No primer for this exon
downstream ENSMUSE00000248738 Chr13:81826125..81826175 TTGGCATCATCTCTCTTGGA Chr13:81826121..81826140 59.32 45
downstream ENSMUSE00000248728 Chr13:81821111..81821193 No primer for this exon
downstream ENSMUSE00000248719 Chr13:81817115..81817258 GATCTTTCACCCTCGCCTCT Chr13:81817205..81817224 60.74 55
downstream ENSMUSE00000374931 Chr13:81812836..81815121 TGGATGTAAGCGTTCAGCAG Chr13:81813474..81813493 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTCCACCCCTGTTTCCTGTA Chr13:81835738..81835758 59.96 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTCCACCCCTGTTTCCTGTA Chr13:81835738..81835758 59.96 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000035834