Method and Apparatus For Isolation of Nucleic Acid Using Lysing Agent Effective in Neutralizing Cellular Contaminants and Sample Contaminants
A method and apparatus to isolate nucleic acids using lysing agent is described that is also effective in neutralizing cellular and sample contaminants. The lysing agent is also effective in neutralizing polyphenolic compounds and humic acid like compounds. One advantage of the described method is improvement in efficiency and speed of the nucleic acid isolation process by increasing yield. The method streamlines the protocol with reduced number of steps which in turn reduces the loss of nucleic acid. The described method also improves the purity of extracted nucleic acids. Although the methods describe a protocol for isolation of nucleic acid, they may be generically extended to downstream applications such as nucleic acid amplification (for example, polymerase chain reaction), digestion by restriction enzymes, and sequencing reactions.
Method and Apparatus for Purification of Contaminated Nucleic Acid Pellets Using Wash Buffer Effective in Neutralizing Cellular and Sample Contaminants
A method and apparatus to purify contaminated nucleic acid pellets using a wash buffer is described that is effective in neutralizing cellular contaminants and sample contaminants such as polyphenolic compounds and humic acid like compounds. The described methods are generic and may be applicable to isolation of RNA, DNA or protein. The current invention is a single step purification of contaminated nucleic acid pellets obtained through precipitation or nucleic acids bound to solid state matrices along with contaminants. Although the methods describe a step in the protocol for isolation of nucleic acids, they may be used as a stand-alone protocol for purifying previously isolated nucleic acids that are contaminated. The method is also extended to downstream applications such as nucleic acid amplification (for example, polymerase chain reaction and qPCR), digestion by restriction enzymes, sequencing reactions, metagenomics sequencing, etc.