Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI1452
Trapped Gene
Lgr4 (ENSMUSG00000050199)
Vector Insertion
Chr 2: 109758463 - 109810848
Public Clones (sanger) (sanger) CD0570 (sanger) (sanger) (sanger) (sanger)
(sanger) LST032 (baygenomics) LST020 (baygenomics) RST357 (baygenomics)
5SD071D12 (ggtc) (ggtc) CMHD-GT_326E9-3 (cmhd) IST10070H3 (tigm)
IST10914H1 (tigm) IST14216B8 (tigm) IST15002H7 (tigm) IST14723E1 (tigm)
Private Clones OST385829 (lexicon) OST357914 (lexicon) OST291563 (lexicon) OST287414 (lexicon)
OST279764 (lexicon) OST176504 (lexicon) OST175783 (lexicon) OST100068 (lexicon)
Severity of mutation (?) Insertion after 6% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000686433 (Chr2:109757804..109758462 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAACCAGGGTGTTTGTGAGA Chr2:109757854..109757873 59.56 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000686433 (Chr2:109757804..109758462 +)
Downstram Exon
ENSMUSE00000376695 (Chr2:109810849..109810920 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAACCAGGGTGTTTGTGAGA Chr2:109757854..109757873 59.56 50 No primer for this exon

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000686433 Chr2:109757804..109758462 CAACCAGGGTGTTTGTGAGA Chr2:109757854..109757873 59.56 50

*** Putative Vector Insertion (Chr 2: 109758463 - 109810848) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000376695 Chr2:109810849..109810920 No primer for this exon
downstream ENSMUSE00000315583 Chr2:109829862..109829933 GGTCGTTACCAGCCAGTTGT Chr2:109829884..109829903 60.03 55
downstream ENSMUSE00000315577 Chr2:109831261..109831332 CAAAGCACTCAGTCCACGAA Chr2:109831327..109831346 60.03 50
downstream ENSMUSE00000315571 Chr2:109836711..109836926 GGGGATGCTTGAGATGTTGT Chr2:109836891..109836910 59.93 50
downstream ENSMUSE00000315564 Chr2:109837750..109837821 No primer for this exon
downstream ENSMUSE00000315555 Chr2:109839563..109839631 GCCTGAGGAAATTCATCCAA Chr2:109839604..109839623 60.01 45
downstream ENSMUSE00000315550 Chr2:109840453..109840524 CTCCATCCGGGATAACAGAA Chr2:109840496..109840515 59.89 50
downstream ENSMUSE00000315544 Chr2:109840717..109840788 GCTGAGTTCCCCACAAAAGA Chr2:109840761..109840780 60.23 50
downstream ENSMUSE00000315536 Chr2:109841087..109841155 No primer for this exon
downstream ENSMUSE00000345648 Chr2:109842623..109842694 TTTTGGCACAGATCATCAGG Chr2:109842676..109842695 59.65 45
downstream ENSMUSE00000440130 Chr2:109843611..109843676 CCAATGCACGACAACCATTA Chr2:109843672..109843691 60.38 45
downstream ENSMUSE00000440122 Chr2:109844708..109844779 GGGATGTTAGGCCTTGAAAA Chr2:109844772..109844791 59.02 45
downstream ENSMUSE00000440118 Chr2:109846661..109846732 AGCTTCGCAAAAGCTCCACT Chr2:109846717..109846736 61.57 50
downstream ENSMUSE00000440113 Chr2:109847302..109847427 TCAGGCCTTCCGTAGGAAAT Chr2:109847351..109847370 60.95 50
downstream ENSMUSE00000359323 Chr2:109848242..109848357 GGTCTTGGGGGCTGTTATCT Chr2:109848339..109848358 60.33 55
downstream ENSMUSE00000315414 Chr2:109848739..109848822 No primer for this exon
downstream ENSMUSE00000476247 Chr2:109851408..109854412 AGCTGTCCGAGACAAAGGAA Chr2:109852589..109852608 59.99 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGCTCCTTGTTGGTTTTCAG Chr2:109809430..109809450 59.71 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTTCCGTGACTGGGAAAACC Chr2:109809510..109809530 62.17 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000050199