Jacobs, "What Are We to Keep?" DttP
: Documents to the People 43, no.1 (Spring 2015): 13-15.
is the largest facility investment our company has made in its 90-year history, and we believe that investing in Texas and keeping jobs in the United States is mutually beneficial for both our company and the economy," said Goodman Global Takeshi Ebisu.
The Invitrogen kit (Life Technologies/USA) was used for the reaction, with 1X PCR Buffer (500mM KCl, 100mM Tris-HCl [pH 9.0]); 5[micro]L (1mM each) dATP, dCTP, dGTP e dTTP
; 0.5[micro]L from each primer, 1.0U Taq DNA polymerase and 2mM Mg[Cl.sub.2].
However, each of these reaction mixtures contained 20 nM of the template, 10 nM of each primer 5' -TATTGATTGTGAATTA(C/G)-3', and 100 [micro]M of a single dNTP (dCTP, dGTP, dATP, or dTTP
The PCR reactions included approximately 100 ng of genomic DNA, 2.5 [micro]l of 10x buffer (Tris-HCl, pH 9.0), PCR enhancers, (N[H.sub.4])2S[O.sub.4], 20 mM Mg[Cl.sub.2]), 2.0 [micro]l of 10 mM dNTPs mixture (2.5 mM each of dATP, dCTP, dGTP and dTTP
), 1 of 10 pmol of each primer and 1 U HS Prime Taq (GeNet Bio, Korea) in a 25 [micro]l reaction volume.
The PCR reaction mixture consisted of 1200 nM of each primer, 200 nM probe, 0.4 U AmpliTaq gold polymerase, 200 nM each of dATP, dCTP, dGTP, and dTTP
, 3.5 mM Mg[Cl.sub.2], and 1 x Taqman buffer A containing a reference dye.
100ng of DNA was dissolved in a 25[micro]l system that included 1.1 [micro]mol/LMg[Cl.sub.2], 200 [micro]mol/ LdNTP(dATP, dCTP, dGTP, dTTP
), 1[micro]mol/L primers, and 1.5uTaq enzyme.
Polymerase chain reactions (PCRs) were performed for each individual in a 50-[micro]l total volume consisting of the following: 5[micro]l of DNA (10 ng/ml) from the individual; 1.5 [micro]l of a 10-mM solution of each primer (P-2 and P-8; Griffiths et al., 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; 0.2 [micro]l of Taq DNA polymerase (Promega: GoTaq Flexi DNA Polymerase; Promega Corp., Madison, Wisconsin); 10 [micro]l of a 5X reaction buffer (Promega); 24.8 [micro]l of sterile water.
A 2 [micro]L volume of DNA template was added to 23 [micro]L of reaction mixture, which contained 2.5 [micro]L of PCR buffer 10x (10 mM Tris HCl, 25 mM KCl), 1mM of Mg[Cl.sub.2], 0.5 [micro]M of each primer, 100 [micro]M of each deoxynucleotide trifosfate (dATP, dGTP, dCTP, dTTP
) and 1.5U of Taq polimerase (Invitrogen--Sao Paulo, Brazil).
Decision Support Systems are often used as online alternatives to the development and analysis of courses of action (ACOA) and as tools that can be used for Online Doctrine and Tactics Techniques, and Procedures (DTTP
) for support to operations.
dATP, dCTP, dTTP
, dGTP--1,25 mM each--were used to amplify intron 1 with Taq DNA polymerase.