Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI22544
Trapped Gene
Usp49 (ENSMUSG00000023984)
Vector Insertion
Chr 17: 47810368 - 47811664
Public Clones not available
Private Clones OST292368 (lexicon) OST237835 (lexicon)
Severity of mutation (?) Insertion after 65% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000136950 (Chr17:47808993..47810367 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GTGCAAGGACTACGTGCTCA Chr17:47809254..47809273 60.06 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000136950 (Chr17:47808993..47810367 +)
Downstram Exon
ENSMUSE00000319461 (Chr17:47811665..47811869 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GTGCAAGGACTACGTGCTCA Chr17:47809254..47809273 60.06 55 AGATCCCAAAAGGGCTCAAT Chr17:47811723..47811742 59.9 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000497657 Chr17:47767639..47767864 CAGGAGTGGGAAACGAATGT Chr17:47767742..47767761 59.97 50
upstream ENSMUSE00000503992 Chr17:47786899..47786972 GGAAACCCAGCCAGTGATAA Chr17:47786912..47786931 59.93 50
upstream ENSMUSE00000136950 Chr17:47808993..47810367 GTGCAAGGACTACGTGCTCA Chr17:47809254..47809273 60.06 55

*** Putative Vector Insertion (Chr 17: 47810368 - 47811664) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000319461 Chr17:47811665..47811869 AGATCCCAAAAGGGCTCAAT Chr17:47811723..47811742 59.9 45
downstream ENSMUSE00000136947 Chr17:47815793..47815901 TTTAAGGTGTAGCCGGAGGA Chr17:47815896..47815915 59.7 50
downstream ENSMUSE00000450395 Chr17:47816613..47816818 GACCACTGCGGAGAGATCAT Chr17:47816757..47816776 60.23 55
downstream ENSMUSE00000319430 Chr17:47817631..47821015 AGGAGTGTCCACGGCATAAC Chr17:47818510..47818529 60 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GTGTGTACCCGAAGCAAGGT Chr17:47810377..47810397 60.03 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GACTCGTGACTGGGAAAACC Chr17:47810415..47810435 59.56 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023984