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Example sentences for: perkin-
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Purified PCR products were sequenced using an ABI PRISM 377 (Perkin-Elmer, Norwalk, CT) through the Oregon State University Center for Gene Research and Biotechnology.
Genomic DNA and total RNA were extracted from fibroblast cultures established from skin biopsies as described [ 14 ] . Long-range RT-PCR amplification of FBN1 cDNA fragments and restriction enzyme digestions were performed as previously reported [ 8 ] . Fragments migrating abnormally in agarose gel electrophoresis were excised, purified with the QIAquick Gel Extraction Kit (QIAGEN) and directly sequenced with fluorescent terminators on an ABI Prism 377 DNA sequencer (Perkin-Elmer).
Aliquots of cDNA target template were diluted serially and mixed with 300 nM of primers and 200 nM of fluorogenic probe (Perkin-Elmer, Foster City, CA).
In general, PCR reactions were performed in a volume of 50 μl with 50 picomoles of each primer, dNTP's (Amersham Pharmacia Biotech, Inc., Piscataway, NJ) at a final concentration of 0.5 mM, 1 μl of template, 1X PCR buffer (10X PCR buffer contained 700 mM Tris pH 8.8, 20 mM MgCl 2 , 1% Triton X-100, and 0.1% Tween 20) and Taq polymerase (Perkin-Elmer Inc., Wellesley MA) diluted to the manufacturers recommendations.
Electroporation of DNA into Y. pestis was done as described previously [ 32 ] . Gene amplification was performed with DeepVent (New England Biolabs, Beverly, MA) or Taq (Eppendorf Scientific, Westbury, NY) DNA polymerase in a Perkin-Elmer GeneAmp Model 2400 thermocycler (Applied Biosystems, Foster City, CA).
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