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0]); 5[micro]L (1mM each) dATP, dCTP, dGTP e dTTP; 0.
The quantification of deoxyribonucleoside triphosphate (dNTP) pools, which are the products of RR metabolism, revealed that EGCG, EA, and RA significantly depleted dATP pools with EA also causing a decrease of dCTP pools.
21) The subsequent injection of dCTP also results in incorporation and light, producing the extended product shown (shaded box, right).
2002); 5[micro]l of a dNTP mixture containing 1 mM each of dATP, dCTP, dGTP, and dTTP; 2[micro]l of a 25-mM solution of magnesium chloride; 0.
0625 mM of each deoxynucleotide triphosphate (dATP, dTTP, dCTP, dGTP), 0.
2], 0, 2 [micro]M de cada oligonucleotideo (G1 e G2), 100 [micro]M de dNTP (dATP, dTTP, dCTP e dGTP) e 2,5U, de Taq DNA Polimerase (Ludiwing [R]).
Polymerase Chain Reactions (PCR) were carried out in a final volume of 25 [micro]l containing 20 ng template DNA, 100[micro]M each dATP, dTTP, dGTP & dCTP, 20 ng of decanucleotide primers, 1.
04 mM of each deoxynucleotide (dTTP, dGTP, dCTP, and dATP); 0.
ABBREVIATIONS: bDNA = Branched DNA; DNA = Deoxyribonucleic Acid; dNTP = Deoxyribonucleotides; dATP = Deoxyadenosine Triphosphate; dCTP = Deoxycytidine Triphosphate; dGTP = Deoxyguanosine Triphosphate; dTTP = Deoxythymidine Triphosphate; FDA = Food and Drug Administration; FRET = Fluorescence Resonance Energy Transfer; MRSA = Methicillin Resistant Staph aureus; MTB = Mycobacterium tuberculosis; NASBA = Nucleic Acid Sequence Based Amplification; PCR = Polymerase Chain Reaction; PFGE = Pulsed-Field Gel Electrophoresis; RT-PCR = Reverse Transcription Polymerase Chain Reaction; Taq = Thermus aquaticus; TMA = Transcription-Mediated Amplification; VRE = Vancomycin Resistant Enterococcus.
Omission of dATP, dTTP, or dGTP during the telomerase extension step of the TRAP reaction resulted in no elongation products being amplified; however, omission of dCTP did not affect the formation of a ladder pattern of telomerase products (data not shown).
The tubes were radioactively labeled by Klenow fragment, using 25 [mu]Ci of [[alpha]-32P] dCTP, 6000 Ci/mmol according to the protocol used by Burow et al.
2], 200 [micron]M de cada dGTP, dATP, dTTP, dCTP (Promega, Madison, WI), O,5 [micron]M de cada iniciador (5'-ATCCAAGACAACACTA CTAA-3' y 5'-AAATATCCTCA AAGTTCC-3') y 2,5 unidades de Taq polimerasa (Corpogen, Colombia).
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