Tools for the biologist enabling optimized use of gene trap clones
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About
UniTrap UNI33232
Trapped Gene
Emilin2 (
ENSMUSG00000024053
)
Vector Insertion
Chr 17: 71605663 - 71624628
Public Clones
IST11824C4 (tigm)
IST11078F12 (tigm)
Private Clones
not available
Severity of mutation
(?)
Insertion after 73% of polypeptide chain
Suggested RT-PCR
(?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000235458 (Chr17:71622714..71624627 -)
Vector Sequence
Forward Primer
Location
Tm
%GC
LacZrt Reverse Primer
Location
CAAATCGCACTCCAGAGACA
Chr17:71622810..71622829
59.98
50
TGGCGAAAGGGGGATGTG
Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000235458 (Chr17:71622714..71624627 -)
Downstram Exon
ENSMUSE00000400126 (Chr17:71605664..71606032 -)
Forward Primer
Location
Tm
%GC
Reverse Primer
Location
Tm
%GC
CAAATCGCACTCCAGAGACA
Chr17:71622810..71622829
59.98
50
TCCATGATGACGCCACTAGA
Chr17:71605764..71605783
60.22
50
Other possible Exon-Primers for RT-PCR
(?)
Exon ID
Exon location
Forward Primer
Primer location
Tm
%GC
upstream
ENSMUSE00000394943
Chr17:71660058..71660326
TTGTGGGGCTCAAGATGTGT
Chr17:71660195..71660214
61.53
50
upstream
ENSMUSE00000235492
Chr17:71659489..71659611
TGCGCCTACATCGTGAATAA
Chr17:71659585..71659604
60.24
45
upstream
ENSMUSE00000235473
Chr17:71630014..71630189
AGAGGACCAGACTGCCAAGA
Chr17:71630088..71630107
59.99
55
upstream
ENSMUSE00000235458
Chr17:71622714..71624627
CAAATCGCACTCCAGAGACA
Chr17:71622810..71622829
59.98
50
upstream
ENSMUSE00000400126
Chr17:71605664..71606032
CATAGCTCGGAAGGCTATGC
Chr17:71605675..71605694
59.97
55
*** Putative Vector Insertion (Chr 17: 71605663 - 71624628) ***
Exon ID
Exon location
Reverse Primer
Primer location
Tm
%GC
downstream
ENSMUSE00000137666
Chr17:71605320..71605352
AGGAGGTGACTTGGGGTAGC
Chr17:71605311..71605330
60.51
60
downstream
ENSMUSE00000137670
Chr17:71604459..71604587
ATACATCCCCATCGTTCACC
Chr17:71604452..71604471
59.49
50
downstream
ENSMUSE00000390252
Chr17:71601516..71602433
GAGACAGACAGGACGGCTTC
Chr17:71602318..71602337
59.99
60
Suggested Genomic-PCR
(?)
Forward
Forward location
Tm
%GC
Reverse name
Reverse
Reverse Location
PCR 1
AACACCTAATCGCCTTGCAG
Chr17:71618563..71618583
60.27
50
L232
GATGTGCTGCAAGGCGATTA
Gene trap vector
PCR 2
GGGTCAGAATCTGGTTGTGC
Chr17:71615617..71615637
60.52
55
LacZ
CCAGGGTTTTCCCAGTCACG
Gene trap vector
Come back to gene
ENSMUSG00000024053