Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI10085
Trapped Gene
Cep68 (
ENSMUSG00000044066
)
Vector Insertion
Chr 11: 20130430 - 20130598
Public Clones
XC757 (baygenomics)
XC755 (baygenomics)
Private Clones
not available
Severity of mutation
(?)
Insertion after 100% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000706151 (Chr11:20130431..20130597 -)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
ACCCTCATCCCTGACAACAG
Chr11:20130468..20130487
59.96
55
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000706151 (Chr11:20130431..20130597 -)
Downstram Exon
ENSMUSE00000440702 (Chr11:20130427..20130597 -)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
ACCCTCATCCCTGACAACAG
Chr11:20130468..20130487
59.96
55
AAGATGATCTGCACGGCTCT
Chr11:20130508..20130527
59.98
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
upstream
ENSMUSE00000370638
Chr11:20138469..20138591
CAGGACCAGCCTTACAGGTC
Chr11:20138495..20138514
59.72
60
upstream
ENSMUSE00000680871
Chr11:20138389..20138591
CAGGACCAGCCTTACAGGTC
Chr11:20138495..20138514
59.72
60
upstream
ENSMUSE00000440688
Chr11:20136009..20136105
TCCCTGACAGAGAGCGTCTT
Chr11:20136056..20136075
60.13
55
upstream
ENSMUSE00000706151
Chr11:20130431..20130597
ACCCTCATCCCTGACAACAG
Chr11:20130468..20130487
59.96
55
*** Putative Vector Insertion (Chr 11: 20130430 - 20130598) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
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
AGCAGTCTGGTGAGCTGGAT
Chr11:20130546..20130566
60.02
55
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
CAAAGCAGTCTGGTGAGCTG
Chr11:20130549..20130569
59.77
55
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000044066