Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24644
Trapped Gene
Rpl24 (ENSMUSG00000022601)
Vector Insertion
Chr 16: 55967284 - 55969622
Public Clones H003C11 (ggtc)
Private Clones OST211537 (lexicon)
Severity of mutation (?) Insertion after 41% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000128658 (Chr16:55967173..55967283 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AAAAGGAATCCTCGGCAGAT Chr16:55967218..55967237 60.04 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000128658 (Chr16:55967173..55967283 +)
Downstram Exon
ENSMUSE00000128665 (Chr16:55969623..55969759 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AAAAGGAATCCTCGGCAGAT Chr16:55967218..55967237 60.04 45 GTGATGGCTCGTTGGAATTT Chr16:55969681..55969700 59.94 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000718751 Chr16:55966456..55966551 TCTCTTCGCCATCTTTACCC Chr16:55966505..55966524 59.27 50
upstream ENSMUSE00000699413 Chr16:55966501..55966551 TCTCTTCGCCATCTTTACCC Chr16:55966505..55966524 59.27 50
upstream ENSMUSE00000364485 Chr16:55966506..55966551 AGGCCGACATCTATCACCAT Chr16:55966529..55966548 59.39 50
upstream ENSMUSE00000223023 Chr16:55966660..55966735 GCAGTTTCAGCGGGTACAAG Chr16:55966671..55966690 60.83 55
upstream ENSMUSE00000128658 Chr16:55967173..55967283 AAAAGGAATCCTCGGCAGAT Chr16:55967218..55967237 60.04 45

*** Putative Vector Insertion (Chr 16: 55967284 - 55969622) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000128665 Chr16:55969623..55969759 GTGATGGCTCGTTGGAATTT Chr16:55969681..55969700 59.94 45
downstream ENSMUSE00000128661 Chr16:55970225..55970288 TGATGCCTGCTTAGCCTTTT Chr16:55970264..55970283 59.98 45
downstream ENSMUSE00000222998 Chr16:55971424..55971547 ACACTAGCGTTTCCCACCAA Chr16:55971510..55971529 60.55 50
downstream ENSMUSE00000713691 Chr16:55980708..55980834 TTTCCGAAGCATCCGATATT Chr16:55980803..55980822 59.51 40

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAAGAAAGGGCAGTCGGTAA Chr16:55967269..55967289 60.24 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAAGAAAGGGCAGTCGGTAA Chr16:55967269..55967289 60.24 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000022601