Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14725
Trapped Gene
Rpl18 (ENSMUSG00000059070)
Vector Insertion
Chr 7: 52973510 - 52974622
Public Clones PST15314-NL (escells) PST6807-NR (escells)
Private Clones OST259263 (lexicon)
Severity of mutation (?) Insertion after 1% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000674278 (Chr7:52973474..52973509 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
No primer for this exon TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000674278 (Chr7:52973474..52973509 +)
Downstram Exon
ENSMUSE00000517867 (Chr7:52974623..52974709 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
No primer for this exon ACCTTTCGGTCCTTGTTGTG Chr7:52974660..52974679 60.01 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000674277 Chr7:52970827..52971212 GGGAGGCTCATGGACTTACA Chr7:52971100..52971119 60.07 55
upstream ENSMUSE00000674276 Chr7:52971518..52973509 GTTTACTCGGTCCTCCCACA Chr7:52973294..52973313 59.97 55
upstream ENSMUSE00000674278 Chr7:52973474..52973509 No primer for this exon

*** Putative Vector Insertion (Chr 7: 52973510 - 52974622) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000517867 Chr7:52974623..52974709 ACCTTTCGGTCCTTGTTGTG Chr7:52974660..52974679 60.01 50
downstream ENSMUSE00000520562 Chr7:52974899..52975006 GTTCGGCTCATGAACAACCT Chr7:52974978..52974997 60.12 50
downstream ENSMUSE00000515956 Chr7:52975290..52975388 AATCCGCACATCATCTGTGA Chr7:52975370..52975389 60.08 45
downstream ENSMUSE00000500647 Chr7:52975664..52975857 CTTGCCAAAATGTCGGTACA Chr7:52975831..52975850 59.58 45
downstream ENSMUSE00000512079 Chr7:52976084..52976204 No primer for this exon
downstream ENSMUSE00000513613 Chr7:52976084..52976200 No primer for this exon

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAATCGCCTTGCAGCACATC Chr7:52973561..52973581 62.25 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CCTCACGTGACTGGGAAAAC Chr7:52973556..52973576 60.54 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000059070