Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI13725
Trapped Gene
Lix1 (ENSMUSG00000047786)
Vector Insertion
Chr 17: 17580753 - 17582931
Public Clones CMHD-GT_269F11-3 (cmhd)
Private Clones OST187048 (lexicon) OST136651 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000391086 (Chr17:17580612..17580752 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GAAGCAGTAGCCTCCACCAG Chr17:17580732..17580751 60.01 60 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000391086 (Chr17:17580612..17580752 +)
Downstram Exon
ENSMUSE00000346882 (Chr17:17582932..17583027 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GAAGCAGTAGCCTCCACCAG Chr17:17580732..17580751 60.01 60 CATCCGCATCATCTAACGTG Chr17:17582956..17582975 60.1 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000358111 Chr17:17539650..17539990 ACTGCATCAATGAGCTGCAC Chr17:17539817..17539836 60.02 50
upstream ENSMUSE00000255754 Chr17:17564120..17564283 GAGCTGCTTTGGCAACTTTC Chr17:17564262..17564281 60.14 50
upstream ENSMUSE00000391086 Chr17:17580612..17580752 GAAGCAGTAGCCTCCACCAG Chr17:17580732..17580751 60.01 60

*** Putative Vector Insertion (Chr 17: 17580753 - 17582931) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000346882 Chr17:17582932..17583027 CATCCGCATCATCTAACGTG Chr17:17582956..17582975 60.1 50
downstream ENSMUSE00000658211 Chr17:17586000..17586101 AGGACTGGCCCACAGAAAAT Chr17:17586032..17586051 60.88 50
downstream ENSMUSE00000555243 Chr17:17591224..17591301 CGAGAGCACTTTGTTTCACG Chr17:17591300..17591319 59.63 50
downstream ENSMUSE00000658210 Chr17:17594070..17596349 GTGGACATTGCCAAACACAG Chr17:17595395..17595414 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTGGCCTCTGTGATAATCG Chr17:17580790..17580810 59.67 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TCTGTGACGTGACTGGGAAA Chr17:17580797..17580817 60.28 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000047786