Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI6372
Trapped Gene
Cep68 (ENSMUSG00000044066)
Vector Insertion
Chr 11: 20138592 - 20139198
Public Clones YHC146 (baygenomics) A006F02 (ggtc) IST10752A6 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 92% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000333483 (Chr11:20139199..20140701 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTCCTCTCCCGAGTTGACTG Chr11:20140371..20140390 59.98 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000333483 (Chr11:20139199..20140701 -)
Downstram Exon
ENSMUSE00000680871 (Chr11:20138389..20138591 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTCCTCTCCCGAGTTGACTG Chr11:20140371..20140390 59.98 60 CATCCCCGTGGATAAACAAG Chr11:20138410..20138429 60.18 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000337123 Chr11:20149310..20149421 No primer for this exon
upstream ENSMUSE00000680880 Chr11:20149310..20149374 No primer for this exon
upstream ENSMUSE00000389471 Chr11:20141889..20142241 GAACCAGATCAGCCCCTACA Chr11:20142079..20142098 60.07 55
upstream ENSMUSE00000712621 Chr11:20141889..20142241 GAACCAGATCAGCCCCTACA Chr11:20142079..20142098 60.07 55
upstream ENSMUSE00000333483 Chr11:20139199..20140701 CTCCTCTCCCGAGTTGACTG Chr11:20140371..20140390 59.98 60

*** Putative Vector Insertion (Chr 11: 20138592 - 20139198) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000370638 Chr11:20138469..20138591 GACCTGTAAGGCTGGTCCTG Chr11:20138473..20138492 59.72 60
downstream ENSMUSE00000680871 Chr11:20138389..20138591 CATCCCCGTGGATAAACAAG Chr11:20138410..20138429 60.18 50
downstream ENSMUSE00000440688 Chr11:20136009..20136105 CTGGGGTGTTATCCAACAGG Chr11:20135987..20136006 60.22 55
downstream ENSMUSE00000706151 Chr11:20130431..20130597 AAGATGATCTGCACGGCTCT Chr11:20130508..20130527 59.98 50
downstream ENSMUSE00000440702 Chr11:20130427..20130597 AAGATGATCTGCACGGCTCT Chr11:20130508..20130527 59.98 50
downstream ENSMUSE00000655325 Chr11:20128509..20129484 AAAGACTCCCCGCTTTTCAT Chr11:20128787..20128806 60.07 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGATGTTTCCCCAGCCTAAT Chr11:20139143..20139163 59.39 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 ACGACAGATGAGCAGGAGAAA Chr11:20139221..20139242 60.01 47.62 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector
PCR 3 CCCCCTTGTTATTGCAGATG Chr11:20140697..20140717 60.32 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 4 CCCCCTTGTTATTGCAGATG Chr11:20140697..20140717 60.32 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000044066