Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25858
Trapped Gene
St3gal2 (ENSMUSG00000031749)
Vector Insertion
Chr 8: 113486263 - 113491316
Public Clones not available
Private Clones OST157317 (lexicon)
Severity of mutation (?) Insertion after 58% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000212665 (Chr8:113486069..113486262 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
ACAACACCAACGAAGTGCTG Chr8:113486094..113486113 59.79 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000212665 (Chr8:113486069..113486262 +)
Downstram Exon
ENSMUSE00000310161 (Chr8:113491317..113491496 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
ACAACACCAACGAAGTGCTG Chr8:113486094..113486113 59.79 50 GCTGGCAATCCACATTAGGT Chr8:113491473..113491492 59.96 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000439990 Chr8:113443822..113444005 ACTGGAGAGCGGTGAAGCTA Chr8:113443860..113443879 60.16 55
upstream ENSMUSE00000310174 Chr8:113480658..113481950 TGGCTTAGAGTCTGGCACCT Chr8:113480852..113480871 60.01 55
upstream ENSMUSE00000212665 Chr8:113486069..113486262 ACAACACCAACGAAGTGCTG Chr8:113486094..113486113 59.79 50

*** Putative Vector Insertion (Chr 8: 113486263 - 113491316) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000310161 Chr8:113491317..113491496 GCTGGCAATCCACATTAGGT Chr8:113491473..113491492 59.96 50
downstream ENSMUSE00000310155 Chr8:113493088..113493133 TTTGTCCACTCGAAGGAAGG Chr8:113493130..113493149 60.22 50
downstream ENSMUSE00000310144 Chr8:113493444..113493563 GGCTGGGTTGTAAATCTGGA Chr8:113493467..113493486 59.93 50
downstream ENSMUSE00000720465 Chr8:113493444..113493723 GGCTGGGTTGTAAATCTGGA Chr8:113493467..113493486 59.93 50
downstream ENSMUSE00000351245 Chr8:113494002..113496380 GAGAAGCACCTTGCATGTGA Chr8:113494667..113494686 59.99 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CCAGAAAGTAATCGCCTTGC Chr8:113486306..113486326 59.85 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GTGAGCTCTCACGGAACAGG Chr8:113486264..113486284 61.01 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000031749