Under physiological conditions, RR maintains stable deoxyribonucleotide
production, which is essential for DNA synthesis; accordingly, inhibition of RR activity eliminates this balance.
The essential components in a standard single target PCR contain target DNA, two oligonucleotide primers, a DNA polymerase, deoxyribonucleotide
triphosphates (dNTPs), MgCl2, KCl, and Tris-HCl buffer.
5 mM magnesium chloride (MgCl2), 200 [micro]M of each deoxyribonucleotide
triphosphates (dNTP), 20 pmol/L of each set of specific primers (metabion international AG, Martinsried, Germany), 1 unit Taq DNA polymerase (MBI Fermentas, Amherst, NY, USA), and 0.
These assays measure the incorporation of radiolabelled deoxyribonucleotide
triphosphates (dNTPs) into mechanically sheared or enzymatically digested complex genomic DNA.
2 [micro]M deoxyribonucleotide
triphosphate mix (Promega, Madison, WI, USA), 3 [micro]M Mg[Cl.
The Austin team's innovative work involves novel methods for developing Deoxyribonucleotide
triphosphates (dNTPs) - the individual building blocks for DNA that are made during Polymerase Chain Reaction (PCR), a process to amplify specific cloned or genomic DNA during genetic sequencing.
5 [micro]L 10 mM deoxyribonucleotide
triphosphate, and 0.
We purchased the Superscript II Reverse Transcriptase (RT) kit, deoxyribonucleotide
triphosphate (dNTPs), and Taq polymerase from Invitrogen (Karlsruhe, Germany); restriction enzymes from New England Biolabs (Frankfurt, Germany); random primers from Roche (Mannheim, Germany); and RNAse inhibitor from Promega (Mannheim, Germany).
Differential polymerase chain reaction (PCR) was performed to assess the GSTM1 polymorphism: 268 base pair (bp) ([beta]-globin) and 215 bp (GSTM1) PCR products were amplified using 30 pmol of primers for the GSTM1 gene and 10 pmol primers for the [beta]-globin gene in a 25 [micro]l reaction (200 [micro]M of each deoxyribonucleotide
The PCR mixture contained 50 ng DNA, 5 [micro]1 2 mM deoxyribonucleotide
triphosphates (dNTPs) (lithium salts), 5 [micro]1 10x Hotstar Buffer, 3 or 5 [micro]1 additional 25 mM MgC[1.
DNA polymerase cannot create links between deoxyribonucleotides
unless a free 3'-OH residue (ribo- or deoxyribonucleotide
) permits the initiation of replication.
PCR amplifications were carried out in 25 [micro]L reaction mixtures containing 2 mM mixture of deoxyribonucleotide
triphosphate phate (dNTP), 2.