Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI674
Trapped Gene
Eef1a1 (ENSMUSG00000037742)
Vector Insertion
Chr 9: 78326666 - 78326764
Public Clones DC0383 (sanger) XE690 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 91% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000531573 (Chr9:78326765..78326999 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGATCGATCGTCGTTCTGGT Chr9:78326877..78326896 59.69 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000531573 (Chr9:78326765..78326999 -)
Downstram Exon
ENSMUSE00000269551 (Chr9:78326265..78326665 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGATCGATCGTCGTTCTGGT Chr9:78326877..78326896 59.69 50 CCGTTCTTCCACCACTGATT Chr9:78326467..78326486 59.97 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000695236 Chr9:78329446..78329478 No primer for this exon
upstream ENSMUSE00000389615 Chr9:78328318..78328495 CAAGCGAACCATCGAAAAGT Chr9:78328338..78328357 60.25 45
upstream ENSMUSE00000531577 Chr9:78328048..78328227 GCGTGAGCGTGGTATCACTA Chr9:78328155..78328174 59.9 55
upstream ENSMUSE00000531576 Chr9:78327659..78327955 TCTGGTTGGAATGGTGACAA Chr9:78327681..78327700 59.94 45
upstream ENSMUSE00000531575 Chr9:78327431..78327581 CCAACTCGTCCAACTGACAA Chr9:78327472..78327491 59.72 50
upstream ENSMUSE00000531574 Chr9:78327085..78327341 TAGACGAGGCAATGTTGCTG Chr9:78327135..78327154 60.01 50
upstream ENSMUSE00000531573 Chr9:78326765..78326999 AGATCGATCGTCGTTCTGGT Chr9:78326877..78326896 59.69 50

*** Putative Vector Insertion (Chr 9: 78326666 - 78326764) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000269551 Chr9:78326265..78326665 CCGTTCTTCCACCACTGATT Chr9:78326467..78326486 59.97 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATAATCGCCTTGCAGCACAT Chr9:78326694..78326714 60.64 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAAGCCCATGTGTGTTGAGA Chr9:78326787..78326807 60.72 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000037742