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The first alkaline lysis was performed by Birnom and Doly in 1979. [1] Since then, slight modifications have been made to the procedure to get to today's most widely used approach. The steps of alkaline lysis can be summarized as the formation of a pellet, resuspension of the pellet in solution, cell lysis, neutralization, and centrifugation.
A 0.5 McFarland standard is prepared by mixing 0.05 mL of 1.175% barium chloride dihydrate (BaCl 2 •2H 2 O), with 9.95 mL of 1% sulfuric acid (H 2 SO 4). [ 1 ] Now there are McFarland standards prepared from suspensions of latex particles, which lengthens the shelf life and stability of the suspensions.
NetPrimer is a gratis web-based tool used for analysing primers used in PCR to amplify a DNA sequence. [2] The software predicts the melting temperature of the primers using the nearest neighbor thermodynamic algorithm.
A lysis buffer is a buffer solution used for the purpose of breaking open cells for use in molecular biology experiments that analyze the labile macromolecules of the cells (e.g. western blot for protein, or for DNA extraction).
OLIGO Primer Analysis Software is a software for DNA primer design. [ 1 ] [ 2 ] The first paper describing this software was published in 1989. [ 3 ] The program is a real time PCR primer and probe search and analysis tool.
For a pure RNA sample, the A 230:260:280 should be around 1:2:1, and for a pure DNA sample, the A 230:260:280 should be around 1:1.8:1. [9] Absorption at 330 nm and higher indicates particulates contaminating the solution, causing scattering of light in the visible range. The value in a pure nucleic acid sample should be zero. [citation needed]
TE buffer is also known as T 10 E 1 buffer, which can be read as "T ten E one buffer". To make a 100 ml solution of T 10 E 1 buffer, 1 ml of 1 M Tris base (pH 10–11) and 0.2 ml EDTA (0.5 M) are mixed and made up with double distilled water up to 100ml.
A random hexamer or random hexonucleotides are for various PCR applications such as rolling circle amplification to prime the DNA.. They are oligonucleotide sequences of 6 bases which are synthesised entirely randomly to give a numerous range of sequences that have the potential to anneal at many random points on a DNA sequence and act as a primer to commence first strand cDNA synthesis.