Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24112
Trapped Gene
Clns1a (ENSMUSG00000025439)
Vector Insertion
Chr 7: 104845380 - 104854098
Public Clones (sanger) (sanger) (sanger) (sanger) (sanger) (sanger)
(sanger) (sanger) CMHD-GT_534F11-5S (cmhd)
Private Clones OST234271 (lexicon)
Severity of mutation (?) Insertion after 19% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000364679 (Chr7:104845212..104845379 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGCAATGAGCTTCCTCAAAA Chr7:104845251..104845270 60.33 45 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000364679 (Chr7:104845212..104845379 +)
Downstram Exon
ENSMUSE00000150854 (Chr7:104854099..104854235 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGCAATGAGCTTCCTCAAAA Chr7:104845251..104845270 60.33 45 TCTTGAGGGTAAGCGTTTGG Chr7:104854203..104854222 60.24 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000364679 Chr7:104845212..104845379 GGCAATGAGCTTCCTCAAAA Chr7:104845251..104845270 60.33 45

*** Putative Vector Insertion (Chr 7: 104845380 - 104854098) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000150854 Chr7:104854099..104854235 TCTTGAGGGTAAGCGTTTGG Chr7:104854203..104854222 60.24 50
downstream ENSMUSE00000150852 Chr7:104860210..104860308 GGGCACAAATCGAAATTCAG Chr7:104860295..104860314 60.45 45
downstream ENSMUSE00000150857 Chr7:104861137..104861244 TCGTCCGAGTCTTCATCCTC Chr7:104861212..104861231 60.35 55
downstream ENSMUSE00000150856 Chr7:104862417..104862590 AATCTCTCCAACGTGGCTTG Chr7:104862510..104862529 60.26 50
downstream ENSMUSE00000150848 Chr7:104864961..104865049 GTTGGTGTGGTTTCCACCTC Chr7:104864991..104865010 60.26 55
downstream ENSMUSE00000387819 Chr7:104866729..104867318 TATTGCGAGATGTGGCTGAG Chr7:104867197..104867216 59.97 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGGTGCTTTTGCTGAGGATT Chr7:104845377..104845397 60.78 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGGTGCTTTTGCTGAGGATT Chr7:104845377..104845397 60.78 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000025439