Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI15310
Trapped Gene
Rpl7a (ENSMUSG00000062647)
Vector Insertion
Chr 2: 26767092 - 26767538
Public Clones PST1878-2 (escells) PST10861-NR (escells) PST1250-1 (escells) PST4686-NL (escells)
Private Clones not available
Severity of mutation (?) Insertion after 34% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000468006 (Chr2:26766942..26767091 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAGATTTAACGCGCTTCGTC Chr2:26766963..26766982 59.99 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000468006 (Chr2:26766942..26767091 +)
Downstram Exon
ENSMUSE00000469045 (Chr2:26767539..26767679 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAGATTTAACGCGCTTCGTC Chr2:26766963..26766982 59.99 50 CTTTGCCAGCAGCTTTCTTC Chr2:26767643..26767662 60.27 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000662952 Chr2:26766332..26766350 No primer for this exon
upstream ENSMUSE00000466990 Chr2:26766526..26766646 GGTCAACCCTCTGTTCGAGA Chr2:26766603..26766622 60.24 55
upstream ENSMUSE00000468006 Chr2:26766942..26767091 GAGATTTAACGCGCTTCGTC Chr2:26766963..26766982 59.99 50

*** Putative Vector Insertion (Chr 2: 26767092 - 26767538) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000469045 Chr2:26767539..26767679 CTTTGCCAGCAGCTTTCTTC Chr2:26767643..26767662 60.27 50
downstream ENSMUSE00000471516 Chr2:26767886..26767965 GGTCTACGTCATGGGCAATC Chr2:26767960..26767979 60.35 55
downstream ENSMUSE00000472522 Chr2:26768191..26768321 CCATCTTTCGACACAGAGCA Chr2:26768230..26768249 59.98 50
downstream ENSMUSE00000473587 Chr2:26768508..26768577 TAGCCAGAGCACCCTTGTCT Chr2:26768533..26768552 60.01 55
downstream ENSMUSE00000662951 Chr2:26768685..26768831 GCAATTCGAGCCACAGACTT Chr2:26768743..26768762 60.41 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGTCCCTCCTGCCATTAACC Chr2:26767043..26767063 60.33 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTACCGTGACTGGGAAAACC Chr2:26767139..26767159 59.83 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000062647