Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI8355
Trapped Gene
Mtap1b (ENSMUSG00000052727)
Vector Insertion
Chr 13: 100197413 - 100199166
Public Clones XL067 (baygenomics) A061F06 (ggtc) A061A07 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 95% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000510765 (Chr13:100199167..100205656 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAAAAGTGCAGAGCCTGGAA Chr13:100200857..100200876 59.99 45 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000510765 (Chr13:100199167..100205656 -)
Downstram Exon
ENSMUSE00000498887 (Chr13:100197174..100197412 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAAAAGTGCAGAGCCTGGAA Chr13:100200857..100200876 59.99 45 GTGGACGACTTGGCTGTCTT Chr13:100197357..100197376 60.31 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000514152 Chr13:100286259..100286495 GAGACCGTAACCGAAGAGCA Chr13:100286291..100286310 60.4 55
upstream ENSMUSE00000519850 Chr13:100278063..100278164 GCAACTTGGACCAGGAACTC Chr13:100278112..100278131 59.7 55
upstream ENSMUSE00000511825 Chr13:100214169..100214251 TCGTTCTGATCAACCCTTCA Chr13:100214190..100214209 59.22 45
upstream ENSMUSE00000471664 Chr13:100211589..100211729 CCATAAACTGCTGGTGCTCA Chr13:100211681..100211700 59.86 50
upstream ENSMUSE00000510765 Chr13:100199167..100205656 AAAAAGTGCAGAGCCTGGAA Chr13:100200857..100200876 59.99 45

*** Putative Vector Insertion (Chr 13: 100197413 - 100199166) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000498887 Chr13:100197174..100197412 GTGGACGACTTGGCTGTCTT Chr13:100197357..100197376 60.31 55
downstream ENSMUSE00000504699 Chr13:100194396..100195916 GGCTGCGGTCTCTACAGTTC Chr13:100195728..100195747 60.02 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCGCTTCTCTGTCTGTCCTT Chr13:100199119..100199139 59.6 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCGCTTCTCTGTCTGTCCTT Chr13:100199119..100199139 59.6 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000052727