Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI27923
Trapped Gene
Gss (ENSMUSG00000027610)
Vector Insertion
Chr 2: 155390686 - 155392538
Public Clones not available
Private Clones OST43779 (lexicon)
Severity of mutation (?) Insertion after 78% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000171095 (Chr2:155392539..155392620 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TTGCTGCTCCTAGCCACTTT Chr2:155392567..155392586 60.15 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000171095 (Chr2:155392539..155392620 -)
Downstram Exon
ENSMUSE00000171089 (Chr2:155390496..155390685 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TTGCTGCTCCTAGCCACTTT Chr2:155392567..155392586 60.15 50 TGTACCATTTCCTCCCCGTA Chr2:155390633..155390652 60.18 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000639694 Chr2:155418430..155418466 No primer for this exon
upstream ENSMUSE00000324830 Chr2:155413178..155413312 AGCTCGAAGAACTGGCAAAG Chr2:155413247..155413266 59.76 50
upstream ENSMUSE00000171100 Chr2:155407624..155407769 GCTGTGCAGATGGACTTCAA Chr2:155407681..155407700 59.99 50
upstream ENSMUSE00000171099 Chr2:155404448..155404523 GGACGACTATACTGCCCGTCT Chr2:155404491..155404511 60.53 57.14
upstream ENSMUSE00000171085 Chr2:155404025..155404164 GGCCTGAATCGCTCAGATTA Chr2:155404133..155404152 60.32 50
upstream ENSMUSE00000171081 Chr2:155403258..155403374 AATAACCCCAGCAAGGGACT Chr2:155403303..155403322 59.83 50
upstream ENSMUSE00000171087 Chr2:155398831..155398911 GTGCCGTAGAGAACGAGCTG Chr2:155398839..155398858 61.14 60
upstream ENSMUSE00000171080 Chr2:155398660..155398737 TCTCTGGACCAAAACCGAAG Chr2:155398669..155398688 60.22 50
upstream ENSMUSE00000171092 Chr2:155397255..155397321 CTTCCGAGATGGCTACATGC Chr2:155397275..155397294 60.76 55
upstream ENSMUSE00000171097 Chr2:155393429..155393623 TGGCTGGGACTAAGAAGGTG Chr2:155393537..155393556 60.25 55
upstream ENSMUSE00000171095 Chr2:155392539..155392620 TTGCTGCTCCTAGCCACTTT Chr2:155392567..155392586 60.15 50

*** Putative Vector Insertion (Chr 2: 155390686 - 155392538) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000171089 Chr2:155390496..155390685 TGTACCATTTCCTCCCCGTA Chr2:155390633..155390652 60.18 50
downstream ENSMUSE00000368457 Chr2:155388923..155389455 AGGCACTAGAACCTGCTGGA Chr2:155388988..155389007 60.01 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TTGTCATAATCGCCTTGCAG Chr2:155392473..155392493 59.83 45 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TTGCTGCTCCTAGCCACTTT Chr2:155392565..155392585 60.15 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000027610