Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI14809
Trapped Gene
Ell (ENSMUSG00000070002)
Vector Insertion
Chr 8: 73109928 - 73114418
Public Clones PST2517-NR (escells) PST0019-NR (escells) PST14354-NR (escells) PSTVU01.O9 (vanderbilt)
Private Clones not available
Severity of mutation (?) Insertion after 78% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000581988 (Chr8:73109453..73109927 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CGACCCAAGAAGAAGTCCAA Chr8:73109813..73109832 60.22 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000581988 (Chr8:73109453..73109927 +)
Downstram Exon
ENSMUSE00000581987 (Chr8:73114419..73114479 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CGACCCAAGAAGAAGTCCAA Chr8:73109813..73109832 60.22 50 GTTCATTGTCACAGGCTCCA Chr8:73114448..73114467 59.68 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000581995 Chr8:73063604..73063740 CTCACCGACAGTGCTTTGAA Chr8:73063693..73063712 60.03 50
upstream ENSMUSE00000581994 Chr8:73096762..73096809 AGACCTTCCATCCGATTTGA Chr8:73096777..73096796 59.48 45
upstream ENSMUSE00000581993 Chr8:73102736..73102857 CTATACCCCAGCCTGACTGC Chr8:73102743..73102762 59.72 60
upstream ENSMUSE00000581992 Chr8:73102992..73103155 GGTCACGGTGTGTGCTACTG Chr8:73103037..73103056 60.23 60
upstream ENSMUSE00000581991 Chr8:73105454..73105719 GAAGGATGGGTTGACACAGG Chr8:73105665..73105684 60.36 55
upstream ENSMUSE00000581990 Chr8:73107771..73107895 CTGCAGGACTGCATGTACAAA Chr8:73107807..73107827 59.92 47.62
upstream ENSMUSE00000581989 Chr8:73109056..73109134 GCCACAGAATGCCACTACAG Chr8:73109068..73109087 59.32 55
upstream ENSMUSE00000581988 Chr8:73109453..73109927 CGACCCAAGAAGAAGTCCAA Chr8:73109813..73109832 60.22 50

*** Putative Vector Insertion (Chr 8: 73109928 - 73114418) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000581987 Chr8:73114419..73114479 GTTCATTGTCACAGGCTCCA Chr8:73114448..73114467 59.68 50
downstream ENSMUSE00000581986 Chr8:73114593..73114764 TTGTAGCTCTGACGCTGCTC Chr8:73114640..73114659 59.49 55
downstream ENSMUSE00000581985 Chr8:73115269..73115313 No primer for this exon
downstream ENSMUSE00000581983 Chr8:73115397..73116755 CCTCTAGGGTCTGGGGACTC Chr8:73116276..73116295 60.07 65

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCGTTGCTATCTGCAAGAGG Chr8:73109921..73109941 60.92 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGAGTGCCACTTGGTTGTTT Chr8:73109930..73109950 59.19 45 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000070002