Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI1506
Trapped Gene
Elovl5 (ENSMUSG00000032349)
Vector Insertion
Chr 9: 77808861 - 77823591
Public Clones CC0725 (sanger) G068D07 (ggtc)
Private Clones not available
Severity of mutation (?) Insertion after 28% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000272601 (Chr9:77808673..77808860 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGTATGGCTGGGACCAAAAT Chr9:77808749..77808768 60.19 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000272601 (Chr9:77808673..77808860 +)
Downstram Exon
ENSMUSE00000272581 (Chr9:77823592..77823669 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGTATGGCTGGGACCAAAAT Chr9:77808749..77808768 60.19 45 TGTTCCCTGGCAGAAAAAGT Chr9:77823645..77823664 59.71 45

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000473563 Chr9:77765172..77765333 No primer for this exon
upstream ENSMUSE00000272620 Chr9:77804948..77805013 TCGATGCGTCACTCAGTACC Chr9:77804966..77804985 59.86 55
upstream ENSMUSE00000272601 Chr9:77808673..77808860 TGTATGGCTGGGACCAAAAT Chr9:77808749..77808768 60.19 45

*** Putative Vector Insertion (Chr 9: 77808861 - 77823591) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000272581 Chr9:77823592..77823669 TGTTCCCTGGCAGAAAAAGT Chr9:77823645..77823664 59.71 45
downstream ENSMUSE00000218959 Chr9:77824555..77824726 TGGTAGCGTGGTGGTAGACA Chr9:77824681..77824700 60.17 55
downstream ENSMUSE00000218961 Chr9:77827632..77827756 TGAGGACATGGATGAAGCTG Chr9:77827676..77827695 59.79 50
downstream ENSMUSE00000218960 Chr9:77829281..77829415 TCGTCTGGATGATTGTCAGC Chr9:77829314..77829333 59.79 50
downstream ENSMUSE00000272557 Chr9:77830472..77832323 GTCAATCCTTTCGCTGCTTC Chr9:77830619..77830638 59.96 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGGCCTTCAAGTTCTGGAGT Chr9:77808868..77808888 60.63 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTGGACTCACCCTGCTGTCT Chr9:77808824..77808844 60.86 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032349