Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI32565
Trapped Gene
Lmna (ENSMUSG00000028063)
Vector Insertion
Chr 3: 88287968 - 88288760
Public Clones IST12278A2 (tigm) IST12278A2 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 80% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000175611 (Chr3:88288537..88288759 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCGGTAGAGGAAGTCGATGA Chr3:88288577..88288596 60.36 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000175611 (Chr3:88288537..88288759 -)
Downstram Exon
ENSMUSE00000175613 (Chr3:88287969..88288076 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCGGTAGAGGAAGTCGATGA Chr3:88288577..88288596 60.36 55 GGGAAGCGATAGGTCATCAA Chr3:88287984..88288003 60.04 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000464930 Chr3:88306670..88307229 CAAAGTGCGTGAGGAGTTCA Chr3:88306686..88306705 60.03 50
upstream ENSMUSE00000721382 Chr3:88306670..88307229 CAAAGTGCGTGAGGAGTTCA Chr3:88306686..88306705 60.03 50
upstream ENSMUSE00000673454 Chr3:88297159..88297234 No primer for this exon
upstream ENSMUSE00000419377 Chr3:88292197..88292353 GACCTCGAGGCTCTTCTCAA Chr3:88292285..88292304 59.68 55
upstream ENSMUSE00000175618 Chr3:88290730..88290855 GGCTACAGACGCTGAAGGAG Chr3:88290763..88290782 60.16 60
upstream ENSMUSE00000175619 Chr3:88290382..88290552 TGGAGATCGATAACGGGAAG Chr3:88290493..88290512 60.03 50
upstream ENSMUSE00000175620 Chr3:88290126..88290251 GGCAGTCTGCTGAGAGGAAC Chr3:88290219..88290238 60.14 60
upstream ENSMUSE00000175616 Chr3:88288849..88289069 GGAGATCCATGCCTATCGAA Chr3:88288874..88288893 60 50
upstream ENSMUSE00000175611 Chr3:88288537..88288759 GCGGTAGAGGAAGTCGATGA Chr3:88288577..88288596 60.36 55
upstream ENSMUSE00000175613 Chr3:88287969..88288076 CCACCGAAGTTCACCCTAAA Chr3:88287988..88288007 59.96 50

*** Putative Vector Insertion (Chr 3: 88287968 - 88288760) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000175612 Chr3:88287749..88287868 GCCTTCCACACCAAGTCAGT Chr3:88287788..88287807 60.16 55
downstream ENSMUSE00000175621 Chr3:88287240..88287335 TCTTCTCCATCCTCGTCGTC Chr3:88287237..88287256 60.35 55
downstream ENSMUSE00000419350 Chr3:88287128..88287335 TCTTCTCCATCCTCGTCGTC Chr3:88287237..88287256 60.35 55
downstream ENSMUSE00000708285 Chr3:88287127..88287335 TCTTCTCCATCCTCGTCGTC Chr3:88287237..88287256 60.35 55
downstream ENSMUSE00000500596 Chr3:88286268..88286534 TGACTAGGTTGTCCCCGAAG Chr3:88286287..88286306 60.1 55
downstream ENSMUSE00000673455 Chr3:88285996..88286022 ACATGATGCTGCAGTTCTGG Chr3:88285976..88285995 59.86 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CTAATCGCCTTGCAGCACAT Chr3:88288690..88288710 61.32 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCCGCAACTACCATGACTAA Chr3:88288786..88288806 58.8 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000028063