Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14006
Trapped Gene
Fibp (ENSMUSG00000024911)
Vector Insertion
Chr 19: 5464432 - 5464924
Public Clones CMHD-GT_156B7-3 (cmhd) CMHD-GT_112E8-3 (cmhd)
Private Clones OST359753 (lexicon) OST346872 (lexicon) OST346726 (lexicon)
Severity of mutation (?) Insertion after 94% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000146179 (Chr19:5464334..5464431 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAGCCAGTATTCAGCGTCAG Chr19:5464390..5464409 59.62 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000146179 (Chr19:5464334..5464431 +)
Downstram Exon
ENSMUSE00000339509 (Chr19:5464925..5465051 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAGCCAGTATTCAGCGTCAG Chr19:5464390..5464409 59.62 55 AATCATGATAGAGGCGCAGAA Chr19:5464994..5465014 59.82 42.86

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000393358 Chr19:5460637..5460778 GACATTTTCGTGGGGAACAC Chr19:5460709..5460728 60.22 50
upstream ENSMUSE00000146173 Chr19:5460961..5461159 GAGTACGCTCCGGAATCTTG Chr19:5460982..5461001 59.84 55
upstream ENSMUSE00000146161 Chr19:5461390..5461516 AAGCTGTCCAAGGGTACCAA Chr19:5461436..5461455 59.59 50
upstream ENSMUSE00000146177 Chr19:5462532..5462632 GCAGCACTTCCTCCTCTCTG Chr19:5462600..5462619 60.28 60
upstream ENSMUSE00000146169 Chr19:5463171..5463325 GATTACGCAGCCATCGTCTT Chr19:5463172..5463191 60.24 50
upstream ENSMUSE00000146171 Chr19:5463604..5463712 CCTCCTGGACCTGCATAAGA Chr19:5463692..5463711 60.21 55
upstream ENSMUSE00000146175 Chr19:5463787..5463850 GGAAAGCTGGGTGTCTTCTCT Chr19:5463809..5463829 59.87 52.38
upstream ENSMUSE00000146165 Chr19:5464129..5464215 ATTTGTGGACCTCGTGGAGA Chr19:5464194..5464213 60.51 50
upstream ENSMUSE00000146179 Chr19:5464334..5464431 CAGCCAGTATTCAGCGTCAG Chr19:5464390..5464409 59.62 55

*** Putative Vector Insertion (Chr 19: 5464432 - 5464924) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000339509 Chr19:5464925..5465051 AATCATGATAGAGGCGCAGAA Chr19:5464994..5465014 59.82 42.86

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTGCTGGCTAACTTCCTTGC Chr19:5464449..5464469 60.15 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTGCTGGCTAACTTCCTTGC Chr19:5464449..5464469 60.15 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000024911