Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI9820
Trapped Gene
Cd59a (ENSMUSG00000032679)
Vector Insertion
Chr 2: 103936085 - 103944327
Public Clones XST184 (baygenomics)
Private Clones OST122403 (lexicon) OST122383 (lexicon)
Severity of mutation (?) Insertion after 0% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000686771 (Chr2:103935994..103936084 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CAGTCACTGGCGATCTGAAA Chr2:103936026..103936045 59.98 50 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000686771 (Chr2:103935994..103936084 +)
Downstram Exon
ENSMUSE00000227061 (Chr2:103944328..103944406 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CAGTCACTGGCGATCTGAAA Chr2:103936026..103936045 59.98 50 TGGAACAGAACACAGCCAGA Chr2:103944406..103944425 60.44 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000686771 Chr2:103935994..103936084 CAGTCACTGGCGATCTGAAA Chr2:103936026..103936045 59.98 50

*** Putative Vector Insertion (Chr 2: 103936085 - 103944327) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000227061 Chr2:103944328..103944406 TGGAACAGAACACAGCCAGA Chr2:103944406..103944425 60.44 50
downstream ENSMUSE00000227056 Chr2:103950897..103951001 ACCGGTTGGAAACAGTGGTA Chr2:103950936..103950955 60.27 50
downstream ENSMUSE00000227053 Chr2:103954132..103955342 TTTTGAGCGTGTCAGAGTGG Chr2:103954738..103954757 60.03 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TAAGCAAGGGCACAGAGTTG Chr2:103942087..103942107 59.07 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TAAGCAAGGGCACAGAGTTG Chr2:103942087..103942107 59.07 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032679