Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI34426
Trapped Gene
OTTMUSG00000003947 (ENSMUSG00000075410)
Vector Insertion
Chr 11: 116515292 - 116518435
Public Clones IST14985G12 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 23% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000720642 (Chr11:116514848..116515291 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TCTCCGAGCTAGAAGGCATC Chr11:116514910..116514929 59.68 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000720642 (Chr11:116514848..116515291 +)
Downstram Exon
ENSMUSE00000669684 (Chr11:116518436..116518621 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TCTCCGAGCTAGAAGGCATC Chr11:116514910..116514929 59.68 55 GGTTCCACTCGGTTGGTAAA Chr11:116518624..116518643 59.83 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000714145 Chr11:116514848..116515291 TCTCCGAGCTAGAAGGCATC Chr11:116514910..116514929 59.68 55
upstream ENSMUSE00000720642 Chr11:116514848..116515291 TCTCCGAGCTAGAAGGCATC Chr11:116514910..116514929 59.68 55

*** Putative Vector Insertion (Chr 11: 116515292 - 116518435) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000669684 Chr11:116518436..116518621 GGTTCCACTCGGTTGGTAAA Chr11:116518624..116518643 59.83 50
downstream ENSMUSE00000669683 Chr11:116518852..116518920 CTGCCGGTAGATTGAAGGTC Chr11:116518923..116518942 59.69 55
downstream ENSMUSE00000669689 Chr11:116518852..116518920 CTGCCGGTAGATTGAAGGTC Chr11:116518923..116518942 59.69 55
downstream ENSMUSE00000669682 Chr11:116520612..116520690 No primer for this exon
downstream ENSMUSE00000669688 Chr11:116520612..116520690 No primer for this exon
downstream ENSMUSE00000645982 Chr11:116521047..116521141 ACCGTTTCTCCCGTTTTCTT Chr11:116521134..116521153 59.98 45
downstream ENSMUSE00000669687 Chr11:116521047..116521141 ACCGTTTCTCCCGTTTTCTT Chr11:116521134..116521153 59.98 45
downstream ENSMUSE00000705776 Chr11:116521079..116521096 No primer for this exon
downstream ENSMUSE00000645981 Chr11:116521099..116521141 ACCGTTTCTCCCGTTTTCTT Chr11:116521134..116521153 59.98 45
downstream ENSMUSE00000645980 Chr11:116529346..116529701 ATGGTAGAGACGCGATGCTT Chr11:116529439..116529458 59.87 50
downstream ENSMUSE00000669686 Chr11:116529346..116529701 ATGGTAGAGACGCGATGCTT Chr11:116529439..116529458 59.87 50
downstream ENSMUSE00000705775 Chr11:116529346..116529605 ATGGTAGAGACGCGATGCTT Chr11:116529439..116529458 59.87 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGCCACTAGATAATCGCCTTG Chr11:116515333..116515354 60.24 47.62 L232 GATGTGCTGCAAGGCGATTA Gene trap vector

Come back to gene ENSMUSG00000075410