Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI25093
Trapped Gene
Dnaja2 (
ENSMUSG00000031701
)
Vector Insertion
Chr 8: 88077310 - 88077815
Public Clones
not available
Private Clones
OST195407 (lexicon)
Severity of mutation
(?)
Insertion after 11% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000212042 (Chr8:88077816..88077875 -)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
GCCAAAGAATACCACCCTGA
Chr8:88077841..88077860
59.93
50
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000212042 (Chr8:88077816..88077875 -)
Downstram Exon
ENSMUSE00000212047 (Chr8:88077086..88077309 -)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
GCCAAAGAATACCACCCTGA
Chr8:88077841..88077860
59.93
50
TCATGTCCTCGCCTCTTCTT
Chr8:88077077..88077096
59.95
50
Other possible Exon-Primers for RT-PCR
(?)
Exon ID
Exon location
Forward Primer
Primer location
Tm
%GC
upstream
ENSMUSE00000400893
Chr8:88079069..88079237
CGAAGCTGTACGACATCCTG
Chr8:88079108..88079127
59.47
55
upstream
ENSMUSE00000212042
Chr8:88077816..88077875
GCCAAAGAATACCACCCTGA
Chr8:88077841..88077860
59.93
50
*** Putative Vector Insertion (Chr 8: 88077310 - 88077815) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
downstream
ENSMUSE00000212047
Chr8:88077086..88077309
TCATGTCCTCGCCTCTTCTT
Chr8:88077077..88077096
59.95
50
downstream
ENSMUSE00000212044
Chr8:88072781..88072861
TTTGGTTGTCTTGCCATTGT
Chr8:88072800..88072819
59.02
40
downstream
ENSMUSE00000212041
Chr8:88072463..88072596
CTGATCATAATGCGCACACC
Chr8:88072503..88072522
60.1
50
downstream
ENSMUSE00000212040
Chr8:88070412..88070608
TTCCCCAGTGAACGTAATCC
Chr8:88070456..88070475
59.79
50
downstream
ENSMUSE00000212039
Chr8:88064207..88064351
CACCACAATCTGACGAGCAT
Chr8:88064216..88064235
59.71
50
downstream
ENSMUSE00000212046
Chr8:88063949..88064076
TCACCTCGAACAACACGAAC
Chr8:88064030..88064049
59.73
50
downstream
ENSMUSE00000212043
Chr8:88062531..88063306
CAAACCCATTGTGTGCAAAG
Chr8:88062629..88062648
60
45
Suggested Genomic-PCR
(?)
Forward
Forward location
Tm
%GC
Reverse name
Reverse
Reverse Location
PCR 1
TCCAAATGCTGGAGACAAAGT
Chr8:88077812..88077833
59.73
42.86
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
TCCAAATGCTGGAGACAAAGT
Chr8:88077812..88077833
59.73
42.86
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000031701