Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI34839
Trapped Gene
Ghrh (ENSMUSG00000027643)
Vector Insertion
Chr 2: 157157514 - 157159216
Public Clones (cmhd) IST14717A3 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 59% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000171437 (Chr2:157159114..157159215 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGTATGCCCGGAAAGTGAT Chr2:157159138..157159157 59.96 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000171437 (Chr2:157159114..157159215 -)
Downstram Exon
ENSMUSE00000171435 (Chr2:157157515..157157605 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGTATGCCCGGAAAGTGAT Chr2:157159138..157159157 59.96 50 ATGCTCTCCAGGGTCATCTG Chr2:157157497..157157516 60.22 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000411777 Chr2:157172308..157172391 No primer for this exon
upstream ENSMUSE00000680658 Chr2:157163092..157163162 GCAGAACCTCAATCGGAGAG Chr2:157163112..157163131 59.95 55
upstream ENSMUSE00000316867 Chr2:157159429..157159528 GGGTGCTCTTTGTGATCCTC Chr2:157159479..157159498 59.66 55
upstream ENSMUSE00000171437 Chr2:157159114..157159215 CTGTATGCCCGGAAAGTGAT Chr2:157159138..157159157 59.96 50
upstream ENSMUSE00000171435 Chr2:157157515..157157605 ACAAGCAGATGACCCTGGAG Chr2:157157524..157157543 60.26 55

*** Putative Vector Insertion (Chr 2: 157157514 - 157159216) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000171434 Chr2:157155233..157155371 CAGAGGACGGAAAAGGTCAG Chr2:157155242..157155261 59.84 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATGCCATCTTCACCACCAAC Chr2:157159177..157159197 60.79 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AAACTCCGTGACTGGGAAAA Chr2:157159151..157159171 59.57 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027643