Journal: Frontiers in Medicine
Article Title: Ability of Polyphosphate and Nucleic Acids to Trigger Blood Clotting: Some Observations and Caveats
Figure Lengend Snippet: Recovery of nucleic acids and polyphosphate (polyP) using various purification methods. Known quantities of bacteriophage lambda DNA (DNA, striped bars), polyG (RNA, white bars), or long-chain polyP (black bars) were processed for nucleic acid purification using Qiaquick, DNeasy, or TRIzol kits, following the manufacturer’s instructions, or with phenol/chloroform extraction. Nucleic acid recovery was quantified using A 260 and polyP recovery by digestion with PPX1 and PiBlue assay. Results are plotted as percent of the input amount in each experiment. For all experiments except the Qiaquick kits, the three samples were: (A) DNA alone (5 µg lambda DNA); (B) RNA alone (5 µg polyG); (C) 5 µg polyP alone; (D) a mixture of 5 µg polyP plus 5 µg of DNA; or (E) 5 µg polyP plus 5 µg of RNA (E) . Experiments with the Qiaquick kit employed 2.5 µg of polyP and/or nucleic acid. Data are mean ± SEM ( n = 3).
Article Snippet: In these experiments, known quantities of purified bacteriophage lambda DNA or a synthetic RNA homopolymer, polyG, either alone or mixed with long-chain polyP, were purified using phenol/chloroform extraction or Qiaquick kits (Qiagen), DNeasy kits (Qiagen), RNeasy Miniprep kits (Qiagen), or TRIzol kits (Thermo Fisher), following the manufacturers’ instructions.
Techniques: Purification, Lambda DNA Preparation, Nucleic Acid Purification