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What is the function of solution 2 in plasmid isolation?
This solution contains the detergent sodium dodecyl sulfate (SDS) which dissolves the cell membrane and denatures proteins. The solution is very alkaline (pH > 12) due to the presence of sodium hydroxide. The high pH aids in denaturing proteins and causes the cleavage of the phosphate bonds in RNA. This eliminates interference from high molecular weight RNA during the plasmid purification. Under highly alkaline conditions, the two strands in non-supercoiled DNA (linear fragments of chromosomal DNA, relaxed and nicked circular DNA) separate and are partially removed from solution. However, this does not occur with supercoiled forms of plasmid DNA because the two strands are intertwined and entangled in a way that prevents them from coming apart. Therefore, supercoiled plasmid remains free in solution.
What is the function of the chromatain?
The chromatain have four major functions. They package DNA into a smaller volume to fit in the cell. They strengthen the DNA to allow mitosis, and they prevent damage to DNA. Chromatain control gene expression and DNA replication.
What are four charactoristics of eubacteria?
eubacteria lack a nucleus, lack histones in their DNA, have no membrane bound organelles, and their DNA is in a circular form.
Describe the function of electricity and the agarose gel in electrophoresis?
The electricity pulls the polar DNA strands through the gel, and shorter DNA strands move farther because they are less inhibited by the gel. The gel acts as drag to separate the different length DNA strands, so different DNA creates specific dye bands.
What is the function of primers?
RNA primers are used to initiate the DNA replication at the template strand. DNA molecules require a free 3' OH, to which it could add the nucleotides. This free 3' OH is provided by the RNA primer. So prior to the synthesis of DNA a short fragment of RNA is synthesized that is later excised and filled with DNA molecules.
