Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI36479
Trapped Gene
Cdc5l (ENSMUSG00000023932)
Vector Insertion
Chr 17: 45545284 - 45547888
Public Clones IST10199C5 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 65% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000136466 (Chr17:45547723..45547887 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CTGAAAATGCCGAGAAGGAA Chr17:45547789..45547808 60.32 45 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000136466 (Chr17:45547723..45547887 -)
Downstram Exon
ENSMUSE00000136463 (Chr17:45545285..45545365 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CTGAAAATGCCGAGAAGGAA Chr17:45547789..45547808 60.32 45 TCCTTTACACGCTCAGCATC Chr17:45545315..45545334 59.03 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000502094 Chr17:45570340..45570686 GCGCCGTTACTACTTTCTCG Chr17:45570427..45570446 60.04 55
upstream ENSMUSE00000136462 Chr17:45564888..45564991 GATTGCCTCATTGCTGCATA Chr17:45564919..45564938 59.8 45
upstream ENSMUSE00000402096 Chr17:45563469..45563630 No primer for this exon
upstream ENSMUSE00000389518 Chr17:45562801..45562928 CCCGATAGACATGGATGAGG Chr17:45562801..45562820 60.29 55
upstream ENSMUSE00000543081 Chr17:45561450..45561549 CGTCTGGCCAATACTCAAGG Chr17:45561498..45561517 60.65 55
upstream ENSMUSE00000136475 Chr17:45556550..45556768 AAGACGAGAGCTTCGAGCAG Chr17:45556728..45556747 60.03 55
upstream ENSMUSE00000367198 Chr17:45553894..45554038 CATCAGCTATCCTCCAAACCA Chr17:45553950..45553970 60.08 47.62
upstream ENSMUSE00000607325 Chr17:45552502..45552690 CAGCATCGCTCTGAGAACAC Chr17:45552537..45552556 59.73 55
upstream ENSMUSE00000136467 Chr17:45550071..45550219 GTGGGCTTAATACCCCACTG Chr17:45550151..45550170 59.31 55
upstream ENSMUSE00000394171 Chr17:45548777..45548939 CTAATGCCACCCCAGGTAGA Chr17:45548858..45548877 59.95 55
upstream ENSMUSE00000136466 Chr17:45547723..45547887 CTGAAAATGCCGAGAAGGAA Chr17:45547789..45547808 60.32 45
upstream ENSMUSE00000136463 Chr17:45545285..45545365 GATGCTGAGCGTGTAAAGGA Chr17:45545337..45545356 59.03 50

*** Putative Vector Insertion (Chr 17: 45545284 - 45547888) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000312815 Chr17:45544745..45544987 TTTCCAGATGGTTCGTAGGG Chr17:45544826..45544845 59.93 50
downstream ENSMUSE00000312798 Chr17:45541543..45541740 CCATGGCTCATTCCTTGTTT Chr17:45541663..45541682 59.93 45
downstream ENSMUSE00000396781 Chr17:45529942..45530154 TTCAAAAGTGCGTAGCTCCA Chr17:45529965..45529984 59.61 45
downstream ENSMUSE00000543072 Chr17:45528841..45529253 ACGGTGCTGGGTAACTTGTC Chr17:45529027..45529046 60.03 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 TGTTAGGCCTCCCCGTAATC Chr17:45547832..45547852 61.19 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CTCCGTTTAGGGTTGTTAGGC Chr17:45547844..45547865 60.01 52.38 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000023932