The protocol is really well-written and elaborate aptamer based...

ARGHYA SETT
ARGHYA SETT
May 15, 2022
protocol Protocol: An Aptamer-based mRNA Affinity Purification Procedure (RaPID) for the Identification of Associated RNAs (RaPID-seq) and Proteins (RaPID-MS) in Yeast
The protocol is really well-written and elaborate aptamer based mRNA purification. Although I have few queries :
1. In the protocol, SC-his media recipe is missing. The exact composition of the selective media is not mentioned in the protocol.
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Rohini R Nair Answered May 19, 2022

Gujarat Biotechnology University

In brief, synthetic medium is prepared by mixing 7 g of synthetic dry mix (Dry mix is prepared by mixing 394 g Yeast nitrogen base with ammonium sulphate with 75 g Amino acid (0.3 g each of arginine, cysteine and proline; 0.45 g each of isoleucine, lysine and tyrosine; 0.75 g each of glutamic acid, phenylalanine and serine; 1.0 g each of aspartate, threonine and valine;) with 850 ml DDW, adding 350 ul of 10 N NaOH, followed by stirring and autoclaving in a large (i.e., 3 liters). Cool to room temperature before use. 100 ml of of 20% glucose (wt/vol) is added, as required, along with 10 ml of a sterile-filtered 100 aminoacid stock solution (see below).

The 100 amino-acid stock is composed of up to six amino acids/bases (e.g., adenine, histidine, leucine, methionine, tryptophan and uracil) for the preparation of specific selective media. For example, SC medium contains all six, whereas synthetic medium lacking uracil (SC-U) would contain all but uracil. The 100 amino-acid stock solution is prepared by first adding 0.4 g of each amino acid/base required to 150 ml DDW, followed by the addition of 3 ml concentrated HCl while stirring, developing the volume up to 200 ml, and then sterile filtering.
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