RNAse destroys the RNA and hence RNAse contamination is a problem in RNA extraction as it breaks down RNA. RNAse enzyme is removed by using RNAse inhibitor or precautions like wearing of gloves, autoclaving tips , using RNAse free water/DEPC treated water is done while performing RTPCR
Proteins are made from the instructions in RNA.
If the DNA is not pure, contaminants include RNA and proteins
The word detergent is used instead of soap in a DNA extraction buffer. Detergent is used to create a hydrophobic environment that favors the precipitation of proteins. Proteins are one of two major contaminants in DNA extraction (the other major contaminant being RNA). When protein precipitate, they can be separated by centrifugation and the DNA isolation procedure can continue.
The sequence of the RNA would be UCG-AUG-UGA.
chloroform is used to denature protein and settle it in the bottom during rna extraction ,also it helps to form organic and inorganic layers in which rna is dissolved in inorganic layer.
Chloroform causes proteins to become denatured and become soluble in the organic phase or interphase, while nucleic acids remain in the aqueous phase.
Chloroform denaturizes the proteins and facilitates the separation of the aqueous and organic phase. If needed the extraction with chloroform is performed two or three times to completely remove the impurities from aqueous layer..
Phenol and chloroform are very hydrophobic and are used to remove non-polar bio- molecules (proteins, fragments of membranes, lipids etc) in the extraction.
The actual role of phenol chloroform isoamyl alcohol in a plasmid DNA extraction is to purify the DNA. The alcohol will act in part as a detergent.
it solubilize the lipids and protein and remove them.
importance of phenol
It solubalize lipids and protiens to remove them from DNA
RNAse destroys the RNA and hence RNAse contamination is a problem in RNA extraction as it breaks down RNA. RNAse enzyme is removed by using RNAse inhibitor or precautions like wearing of gloves, autoclaving tips , using RNAse free water/DEPC treated water is done while performing RTPCR
Proteins, RNA, lipids
to precipitate protein.
DNA extraction is done by three methods: * Organic extraction * inorganic extraction * solid state method In organic extraction, phenol and chloroform are used to create on organic phase in which cells are lysed and DNA is freed. The DNA remains in the aqueous phase. Ethyl alcohol is used to precipitate the DNA. In the inroganic methos, NaCl and EDTA are used for cell lysis. Following this, an approach similar to the organic method is followed. In solid state extraction, DNA is first precipitated in the presence of high slat and low pH conditions. The precipitated DNA is then adsorbed on to a filter membrane surface.