Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI24031
Trapped Gene
Ube2n (
ENSMUSG00000074781
)
Vector Insertion
Chr 10: 95004413 - 95004905
Public Clones
not available
Private Clones
OST237136 (lexicon)
OST15957 (lexicon)
Severity of mutation
(?)
Insertion after 92% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000640784 (Chr10:95004272..95004412 +)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
CAAATGATGTAGCCGAGCAA
Chr10:95004359..95004378
59.83
45
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000640784 (Chr10:95004272..95004412 +)
Downstram Exon
ENSMUSE00000640783 (Chr10:95004906..95008292 +)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
CAAATGATGTAGCCGAGCAA
Chr10:95004359..95004378
59.83
45
GCAGGAGGACTCAAAACTGC
Chr10:95006138..95006157
60
55
Other possible Exon-Primers for RT-PCR
(?)
Exon ID
Exon location
Forward Primer
Primer location
Tm
%GC
upstream
ENSMUSE00000640786
Chr10:95003783..95003815
No primer for this exon
upstream
ENSMUSE00000640785
Chr10:95003839..95004085
CCAATGGCAGCACCTAAAGT
Chr10:95003995..95004014
60.13
50
upstream
ENSMUSE00000640784
Chr10:95004272..95004412
CAAATGATGTAGCCGAGCAA
Chr10:95004359..95004378
59.83
45
*** Putative Vector Insertion (Chr 10: 95004413 - 95004905) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
downstream
ENSMUSE00000640783
Chr10:95004906..95008292
GCAGGAGGACTCAAAACTGC
Chr10:95006138..95006157
60
55
Suggested Genomic-PCR
(?)
Forward
Forward location
Tm
%GC
Reverse name
Reverse
Reverse Location
PCR 1
GTAGCCGAGCAATGGAAGAC
Chr10:95004368..95004388
59.84
55
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
GTAGCCGAGCAATGGAAGAC
Chr10:95004368..95004388
59.84
55
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000074781