Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI23097
Trapped Gene
Ccnb2 (ENSMUSG00000032218)
Vector Insertion
Chr 9: 70266715 - 70266807
Public Clones CMHD-GT_487G10-3 (cmhd)
Private Clones OST276449 (lexicon) OST163956 (lexicon)
Severity of mutation (?) Insertion after 13% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000217822 (Chr9:70266808..70266936 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCGGGCAGTTTTAGAAGAAA Chr9:70266849..70266868 59.47 45 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000217822 (Chr9:70266808..70266936 -)
Downstram Exon
ENSMUSE00000217820 (Chr9:70266601..70266714 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCGGGCAGTTTTAGAAGAAA Chr9:70266849..70266868 59.47 45 CTGCACTGGTTTCACAGAGG Chr9:70266600..70266619 59.46 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000406066 Chr9:70269224..70269354 No primer for this exon
upstream ENSMUSE00000217822 Chr9:70266808..70266936 GCGGGCAGTTTTAGAAGAAA Chr9:70266849..70266868 59.47 45

*** Putative Vector Insertion (Chr 9: 70266715 - 70266807) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000217820 Chr9:70266601..70266714 CTGCACTGGTTTCACAGAGG Chr9:70266600..70266619 59.46 55
downstream ENSMUSE00000217823 Chr9:70260909..70261079 GCAGAGGCTCTCTTCCTTCA Chr9:70261007..70261026 59.83 55
downstream ENSMUSE00000217818 Chr9:70260649..70260807 CAGAAGCCGAAACTTGGAAT Chr9:70260675..70260694 59.31 45
downstream ENSMUSE00000217821 Chr9:70258462..70258698 TTTCTCGGATTTGGGAACTG Chr9:70258529..70258548 60.04 45
downstream ENSMUSE00000217829 Chr9:70257928..70258068 AGCGTCAGCTCCATCAGGTA Chr9:70257997..70258016 60.97 55
downstream ENSMUSE00000318721 Chr9:70257087..70257197 GCTGCATGACTTCCAGGACT Chr9:70257115..70257134 60.42 55
downstream ENSMUSE00000217819 Chr9:70255499..70255836 GAGGGATCGTGCTGATCTTC Chr9:70255760..70255779 59.77 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAAGGACAAGATGGCATGTG Chr9:70266782..70266802 60.11 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CAAGGACAAGATGGCATGTG Chr9:70266782..70266802 60.11 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032218