Hi all,
in your opinion is it a problem that I have an EDTA solution at 9.5 instead of 8 for plasmidic extraction? I added more NaOH to dissolve the powder because at Ph 8 there still were not dissolved residuals.
Do you know some tricks that help the EDTA to dissolve better?
Hi, in my experience I need hours to make an EDTA stock solution 0,5M at pH8 without undissolved salt. Here a discussion that can help you: https://www.researchgate.net/post/I_have_been_preparing_05M_EDTA_ph_8_since_yesterday_and_it_is_not_dissolving_I_need_help_coz_I_seriously_need_to_prepare_TAE_buffer/1
Used a lot of NaOH (I used pellet but I preferred NaOH 2-3N solution), using the disodium salt, magnetic stirring and a lot of patience. When you are near pH8 the salt undissolved began to dissolve and the pH decrease, for this reason you need a lot of patience but when you are around pH8 leave the solution stirring for some time, then adjust pH if needed.
Fot the plasmid question I always used EDTA around pH8 so basically the lysis buffer is around the same pH, so the question is if you can have a lysis buffer more alkaline. I find this protocols for handmade lysis buffer for plasmid extraction from bacteria, https://www.dsmz.de/fileadmin/Bereiche/Microbiology/Dateien/Kultivierungshinweise/Plasmid_Isolation_from_Bacteria.pdf
Some protocols use pH above 8 so technically I think that you can use EDTA, I mean for lysis buffer, at pH 9,5
Hope this can help you
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