Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI38353
Trapped Gene
Gli2 (ENSMUSG00000048402)
Vector Insertion
Chr 1: 120764555 - 120786979
Public Clones (sanger)
Private Clones not available
Severity of mutation (?) Insertion after 5% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000475590 (Chr1:120786873..120786978 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTTTGCCGATTGACATGAGA Chr1:120786926..120786945 59.8 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000475590 (Chr1:120786873..120786978 -)
Downstram Exon
ENSMUSE00000234051 (Chr1:120764556..120764758 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTTTGCCGATTGACATGAGA Chr1:120786926..120786945 59.8 45 CATATGGGGGTTCACGTAGG Chr1:120764613..120764632 60.07 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000692693 Chr1:120949879..120950196 CCTGCATGCTAGAGGCAAAC Chr1:120949892..120949911 60.93 55
upstream ENSMUSE00000692692 Chr1:120898617..120898783 CTCTTGGAGGACAGCAGCTT Chr1:120898679..120898698 59.74 55
upstream ENSMUSE00000475590 Chr1:120786873..120786978 CTTTGCCGATTGACATGAGA Chr1:120786926..120786945 59.8 45
upstream ENSMUSE00000234051 Chr1:120764556..120764758 AGCACTACCTCCGGTCTGTG Chr1:120764614..120764633 60.32 60

*** Putative Vector Insertion (Chr 1: 120764555 - 120786979) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000234129 Chr1:120763149..120763334 CATGAGGGTCATCTGGTGGT Chr1:120763221..120763240 60.81 55
downstream ENSMUSE00000426396 Chr1:120752013..120752214 CTACCAGCGAGTTGGGAGAG Chr1:120752064..120752083 60.01 60
downstream ENSMUSE00000597455 Chr1:120751018..120751234 TGGTAGGCCACAGGATTGAT Chr1:120751168..120751187 60.34 50
downstream ENSMUSE00000234014 Chr1:120749835..120749957 CTGACTCGCTGTTCTGCTTG Chr1:120749914..120749933 59.92 55
downstream ENSMUSE00000133480 Chr1:120744960..120745094 CAGTGGCAGTTGGTCTCGTA Chr1:120744984..120745003 59.9 55
downstream ENSMUSE00000659629 Chr1:120742494..120742643 GGCACACGAACTCCTTCTTC Chr1:120742573..120742592 59.85 55
downstream ENSMUSE00000659628 Chr1:120740928..120741092 GTGTGAACGCAGGTGTGTCT Chr1:120741008..120741027 59.79 55
downstream ENSMUSE00000133483 Chr1:120738493..120738774 GGACATGCACATCATTACGC Chr1:120738620..120738639 59.96 50
downstream ENSMUSE00000133482 Chr1:120736831..120737167 TCCATGCCACTGTCATTGTT Chr1:120737057..120737076 59.97 45
downstream ENSMUSE00000469773 Chr1:120730638..120734754 GGATACGCACATGTCTGTGG Chr1:120731127..120731146 59.99 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGTTCTCTCTGACCCCTGTG Chr1:120765985..120766005 59.68 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GGCCTCCGTTTTATGACCTC Chr1:120766003..120766023 60.83 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000048402