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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.
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.
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.
Add 100 μL of each of the above to separate tubes (use microcentrifuge tubes) and add 1.0 mL of Coomassie Blue to each tube. Turn on and adjust a spectrophotometer to a wavelength of 595 nm, and blank the spectrophotometer using 1.5 mL cuvettes or use a mobile smartphone camera ( RGBradford method).
The annealing temperature during a polymerase chain reaction determines the specificity of primer annealing. The melting point of the primer sets the upper limit on annealing temperature. At temperatures just below this point, only very specific base pairing between the primer and the template will occur.
Primer Premier is a bioinformatics software used for various PCR applications. It supports the design of degenerate primers for amplifying a related set of nucleotide sequences for the detection of common traits amongst organisms, as well as to determine heredity. [1] The software also designs tagged and nested primers for multiplex PCR ...
Loop-mediated isothermal amplification (LAMP) primers [1] Loop-mediated isothermal amplification (LAMP) product [1]. In LAMP, the target sequence is amplified at a constant temperature of 60–65 °C (140–149 °F) using either two or three sets of primers and a polymerase like Bst Klenow fragment with high strand displacement activity in addition to a replication activity.
Primer extension is a technique whereby the 5' ends of RNA can be mapped - that is, they can be sequenced and properly identified. Primer extension can be used to determine the start site of transcription (the end site cannot be determined by this method) by which its sequence is known.