Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI7930
Trapped Gene
Fbxl17 (ENSMUSG00000023965)
Vector Insertion
Chr 17: 63842103 - 63848373
Public Clones RRM165 (baygenomics)
Private Clones not available
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000385763 (Chr17:63848374..63848649 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CCGAGATCCCAGACATCAAC Chr17:63848401..63848420 60.47 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000385763 (Chr17:63848374..63848649 -)
Downstram Exon
ENSMUSE00000693382 (Chr17:63842038..63842102 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CCGAGATCCCAGACATCAAC Chr17:63848401..63848420 60.47 55 ACCAAGGGCTGTAGGTCTCC Chr17:63842025..63842044 60.51 60

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000385763 Chr17:63848374..63848649 CCGAGATCCCAGACATCAAC Chr17:63848401..63848420 60.47 55

*** Putative Vector Insertion (Chr 17: 63842103 - 63848373) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000693382 Chr17:63842038..63842102 ACCAAGGGCTGTAGGTCTCC Chr17:63842025..63842044 60.51 60
downstream ENSMUSE00000308088 Chr17:63840114..63840236 TCACGCCAGTACTTGCAAAC Chr17:63840147..63840166 59.91 50
downstream ENSMUSE00000136693 Chr17:63837061..63837318 CTGGTTGCCTACGTGGACTT Chr17:63837069..63837088 60.17 55
downstream ENSMUSE00000136695 Chr17:63820757..63820888 TGGCCAAAATGGATGTCTTT Chr17:63820823..63820842 60.31 40
downstream ENSMUSE00000693381 Chr17:63800931..63801014 TATCCAGCAGCACAAAACCA Chr17:63800962..63800981 60.26 45
downstream ENSMUSE00000136696 Chr17:63734294..63734401 GAACAACCCATGAAGCCAAC Chr17:63734306..63734325 60.36 50
downstream ENSMUSE00000136692 Chr17:63706097..63706227 ACGGAGGTCTAAGCTGGACA Chr17:63706176..63706195 59.87 55
downstream ENSMUSE00000307985 Chr17:63574374..63574450 TCCTTCCTTTGCAATGACCT Chr17:63574401..63574420 59.67 45
downstream ENSMUSE00000358045 Chr17:63429770..63429912 ATCCGACGTCCACAGTCTCT Chr17:63429837..63429856 59.71 55
downstream ENSMUSE00000439569 Chr17:63407131..63409760 ATATTGCCAGCTGGTTTTGG Chr17:63407755..63407774 59.96 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AACCGGTGGGAAATCTTAGC Chr17:63848366..63848386 60.32 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 AACCGGTGGGAAATCTTAGC Chr17:63848366..63848386 60.32 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023965