Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI5091
Trapped Gene
Cep68 (
ENSMUSG00000044066
)
Vector Insertion
Chr 11: 20130598 - 20136008
Public Clones
(sanger)
(sanger)
YTC650 (baygenomics)
XC762 (baygenomics)
(ggtc)
IST11991F10 (tigm)
IST12720E6 (tigm)
IST10231F2 (tigm)
IST12355F11 (tigm)
IST11551C6 (tigm)
Private Clones
not available
Severity of mutation
(?)
Insertion after 100% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000440688 (Chr11:20136009..20136105 -)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
TCCCTGACAGAGAGCGTCTT
Chr11:20136056..20136075
60.13
55
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000440688 (Chr11:20136009..20136105 -)
Downstram Exon
ENSMUSE00000706151 (Chr11:20130431..20130597 -)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
TCCCTGACAGAGAGCGTCTT
Chr11:20136056..20136075
60.13
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
*** Putative Vector Insertion (Chr 11: 20130598 - 20136008) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
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
TGTTGGATAACACCCCAGGT
Chr11:20133005..20133025
60.09
50
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
AGGCCTGTTTTGAACGTGAC
Chr11:20132951..20132971
60.16
50
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
PCR 3
TCTTAATCGCCTTGCAGCAC
Chr11:20133038..20133058
61.44
50
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 4
TACACGTGACTGGGAAAACC
Chr11:20133039..20133059
58.47
50
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000044066