Further queries as follows:-In procedure. step no 6. the shaking...

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
Further queries as follows:
-In procedure. step no 6. the shaking speed(how much RPM) to induce the expression of the MS2-CP-GFP-SBP is not mentioned. I think the shaking speed is a critical parameter for protein expression.
-For western blot, which primary/secondary antibody sho
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Rohini R Nair Answered May 19, 2022

Gujarat Biotechnology University

-In procedure. step no 6. the shaking speed(how much RPM) to induce the expression of the MS2-CP-GFP-SBP is not mentioned. I think the shaking speed is a critical parameter for protein expression.
170rpm
-For western blot, which primary/secondary antibody should we use to determine MS2-CP-GFP-SBP on the SDS gel. or fluorophore tagged MS2 aptamer can be used to visualize the pull-down fractions?
anti-GFP
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