Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI17381
Trapped Gene
Urm1 (ENSMUSG00000069020)
Vector Insertion
Chr 2: 29697004 - 29698240
Public Clones CMHD-GT_536G7-3 (cmhd)
Private Clones OST443702 (lexicon) OST318462 (lexicon) OST272909 (lexicon) OST265314 (lexicon)
OST253941 (lexicon) OST251879 (lexicon) OST236441 (lexicon) OST233075 (lexicon)
OST220627 (lexicon) OST209058 (lexicon) OST123714 (lexicon) OST109692 (lexicon)
OST69155 (lexicon) OST42234 (lexicon) OST42220 (lexicon) OST40117 (lexicon)
Severity of mutation (?) Insertion after 63% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000568261 (Chr2:29696922..29697003 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGCGACCAGAGCTGTTCATC Chr2:29696970..29696989 60.56 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000568261 (Chr2:29696922..29697003 +)
Downstram Exon
ENSMUSE00000568260 (Chr2:29698241..29698289 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGCGACCAGAGCTGTTCATC Chr2:29696970..29696989 60.56 55 GTTCCCAGTCGGCATCATTA Chr2:29698287..29698306 60.86 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000568263 Chr2:29682904..29682955 CGCCCTTATGTGTGAAGGTG Chr2:29682928..29682947 61.48 55
upstream ENSMUSE00000568262 Chr2:29688212..29688282 GCTCCTGTTTGATGGGGTAA Chr2:29688224..29688243 59.93 50
upstream ENSMUSE00000568261 Chr2:29696922..29697003 AGCGACCAGAGCTGTTCATC Chr2:29696970..29696989 60.56 55

*** Putative Vector Insertion (Chr 2: 29697004 - 29698240) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000568260 Chr2:29698241..29698289 GTTCCCAGTCGGCATCATTA Chr2:29698287..29698306 60.86 50
downstream ENSMUSE00000568259 Chr2:29698623..29699582 GAGTTGTAGGCAGCCTGGAG Chr2:29699372..29699391 60.01 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCCAGGGAGACAGTGTGTGA Chr2:29696989..29697009 60.29 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGTCGTGACTGGGAAAACC Chr2:29697051..29697071 60.54 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000069020