Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI26033
Trapped Gene
Sec61g (ENSMUSG00000078974)
Vector Insertion
Chr 11: 16404842 - 16406375
Public Clones not available
Private Clones OST144132 (lexicon)
Severity of mutation (?) Insertion after 46% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000681063 (Chr11:16406376..16406475 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGACTCAATTCGGCTGGTTA Chr11:16406403..16406422 60.07 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000681063 (Chr11:16406376..16406475 -)
Downstram Exon
ENSMUSE00000681061 (Chr11:16404739..16404841 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGACTCAATTCGGCTGGTTA Chr11:16406403..16406422 60.07 50 TTTCACGAAGAAGCCAATGA Chr11:16404749..16404768 59.4 40

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000681064 Chr11:16408107..16408487 AGCTGTAGCGCAGACACTCA Chr11:16408440..16408459 59.95 55
upstream ENSMUSE00000681058 Chr11:16408103..16408169 CTCAATCCGCCATCCAGTAA Chr11:16408103..16408122 60.99 50
upstream ENSMUSE00000681056 Chr11:16407983..16408072 CAGGGGAGGTGAAGAAACTG Chr11:16407986..16408005 59.69 55
upstream ENSMUSE00000681063 Chr11:16406376..16406475 GGACTCAATTCGGCTGGTTA Chr11:16406403..16406422 60.07 50

*** Putative Vector Insertion (Chr 11: 16404842 - 16406375) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000681061 Chr11:16404739..16404841 TTTCACGAAGAAGCCAATGA Chr11:16404749..16404768 59.4 40
downstream ENSMUSE00000681055 Chr11:16401654..16401810 AGCATTCAGGCAACGCTTTA Chr11:16401708..16401727 60.9 45
downstream ENSMUSE00000681057 Chr11:16401390..16401810 TTCATGGCTCTCAGGATGTG Chr11:16401412..16401431 59.79 50
downstream ENSMUSE00000681059 Chr11:16400533..16401810 GGGAAAGACGTTCACTTCCA Chr11:16400980..16400999 60.09 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TCTCTGCAGTAATCGCCTTG Chr11:16406313..16406333 59.17 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AAATCTCTGCAGCGTGACTG Chr11:16406316..16406336 59.19 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000078974