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Hello,I am about to use your protocol for some RNA purification...

Laura Weise Jul 29, 2020

Protocol: Purification of RNA Mango Tagged Native RNA-protein Complexes from Cellular Extracts Using TO1-Desthiobiotin Fluorophore Ligand

Hello,
I am about to use your protocol for some RNA purification and I wondered about how you are preparing the 10x Biotin elution buffer with 200 mM Biotin in 10x Binding buffer in detail? I am always facing problems to dissolve the biotin in water. Do you dilute the biotin first in Read more more

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Shanker Shyam Sundhar Panchapakesan Author Answered Jul 30, 2020

Molecular, Cellular and Developmental Biology, Yale University, USA, USA,

Sorry, read that as 200 mM biotin stock

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Shanker Shyam Sundhar Panchapakesan Author Answered Jul 30, 2020

Molecular, Cellular and Developmental Biology, Yale University, USA, USA,

Hi Laura,

Solubility of Biotin is very low in water. You can make 100 mM stock or so by titrating with NaOH. Just add biotin in water and Keep on adding NaOH, say from a 2 M stock, till biotin dissolves. You can then use this stock to dilute into elution buffer of your choice. My email is shankershyams.panchapakesan@yale.edu. If you have any questions, you can contact me directly in that email.

Cheers,
Shyam

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