Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI20296
Trapped Gene
A630054L15Rik (ENSMUSG00000040818)
Vector Insertion
Chr 14: 27427699 - 27428021
Public Clones IST13044F6 (tigm)
Private Clones OST368010 (lexicon) OST249170 (lexicon) OST241593 (lexicon) OST170435 (lexicon)
Severity of mutation (?) Insertion after 51% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000251504 (Chr14:27427576..27427698 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
AGATGCTCTGGGAGTTGGTG Chr14:27427605..27427624 60.26 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000251504 (Chr14:27427576..27427698 +)
Downstram Exon
ENSMUSE00000251540 (Chr14:27428022..27428117 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
AGATGCTCTGGGAGTTGGTG Chr14:27427605..27427624 60.26 55 ATCGTGAAGTAAGGCCGAAA Chr14:27428075..27428094 59.71 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000406003 Chr14:27399064..27399348 GCTACACTGCGTGTGTGTGG Chr14:27399294..27399313 61.44 60
upstream ENSMUSE00000355312 Chr14:27411719..27411757 No primer for this exon
upstream ENSMUSE00000398626 Chr14:27411881..27411923 No primer for this exon
upstream ENSMUSE00000251520 Chr14:27419102..27419214 TCTGGGAGAAGGGTGTCACT Chr14:27419146..27419165 59.68 55
upstream ENSMUSE00000251514 Chr14:27419930..27420010 No primer for this exon
upstream ENSMUSE00000251572 Chr14:27422534..27422639 No primer for this exon
upstream ENSMUSE00000370900 Chr14:27424160..27424239 CACCTGCCTATCATGGGACT Chr14:27424210..27424229 59.95 55
upstream ENSMUSE00000398486 Chr14:27425362..27425424 No primer for this exon
upstream ENSMUSE00000353400 Chr14:27426391..27426446 TGCCTACTGTTCACGAGGTG Chr14:27426416..27426435 59.9 55
upstream ENSMUSE00000251504 Chr14:27427576..27427698 AGATGCTCTGGGAGTTGGTG Chr14:27427605..27427624 60.26 55

*** Putative Vector Insertion (Chr 14: 27427699 - 27428021) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000251540 Chr14:27428022..27428117 ATCGTGAAGTAAGGCCGAAA Chr14:27428075..27428094 59.71 45
downstream ENSMUSE00000251495 Chr14:27431453..27431547 CGAATAATGTGTGGCCAGTG Chr14:27431527..27431546 59.99 50
downstream ENSMUSE00000251438 Chr14:27436077..27436142 No primer for this exon
downstream ENSMUSE00000251479 Chr14:27436380..27436444 No primer for this exon
downstream ENSMUSE00000251474 Chr14:27438801..27438890 TAGCGCCGGAGAATAACACT Chr14:27438856..27438875 59.87 50
downstream ENSMUSE00000251469 Chr14:27446192..27446245 CTTTTCTGCAAAGGCATCAAG Chr14:27446231..27446251 60 42.86
downstream ENSMUSE00000251463 Chr14:27446351..27446460 CAGTCGCCCTTTATTCCAGA Chr14:27446448..27446467 60.21 50
downstream ENSMUSE00000251455 Chr14:27446805..27446907 TCTTGAACCAGCCATCAAAA Chr14:27446848..27446867 59.25 40
downstream ENSMUSE00000251448 Chr14:27448158..27448232 CAGCTTCAACACAAGGTCCA Chr14:27448223..27448242 59.87 50
downstream ENSMUSE00000434196 Chr14:27448914..27452880 GGCCTCAGCACTGAACTAGG Chr14:27451413..27451432 60.01 60

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGAGTGCATTTGCCTCTTGA Chr14:27427701..27427721 60.54 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGAGTGCATTTGCCTCTTGA Chr14:27427701..27427721 60.54 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000040818