Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI8759
Trapped Gene
RP23-117O11.3 (
ENSMUSG00000035399
)
Vector Insertion
Chr 2: 163237247 - 163241270
Public Clones
XN859 (baygenomics)
Private Clones
not available
Severity of mutation
(?)
Insertion after 9% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000329327 (Chr2:163241271..163241391 -)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
TTAACGGGAGGGAACATCAA
Chr2:163241370..163241389
60.3
45
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000329327 (Chr2:163241271..163241391 -)
Downstram Exon
ENSMUSE00000329324 (Chr2:163237133..163237246 -)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
TTAACGGGAGGGAACATCAA
Chr2:163241370..163241389
60.3
45
TATCATCGGCTGCTGTTCTG
Chr2:163237154..163237173
59.97
50
Other possible Exon-Primers for RT-PCR
(?)
Exon ID
Exon location
Forward Primer
Primer location
Tm
%GC
upstream
ENSMUSE00000407125
Chr2:163245148..163245206
No primer for this exon
upstream
ENSMUSE00000329327
Chr2:163241271..163241391
TTAACGGGAGGGAACATCAA
Chr2:163241370..163241389
60.3
45
*** Putative Vector Insertion (Chr 2: 163237247 - 163241270) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
downstream
ENSMUSE00000329324
Chr2:163237133..163237246
TATCATCGGCTGCTGTTCTG
Chr2:163237154..163237173
59.97
50
downstream
ENSMUSE00000365176
Chr2:163231561..163232825
TCTTGCTGCCACATCGTAAG
Chr2:163231789..163231808
60.01
50
Suggested Genomic-PCR
(?)
Forward
Forward location
Tm
%GC
Reverse name
Reverse
Reverse Location
PCR 1
GTGGACGCATCAGGGTAACT
Chr2:163241263..163241283
60
55
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
AGAGTGGACGCATCAGGGTA
Chr2:163241266..163241286
60.68
55
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000035399