Re: [DIYbio] Re: Miniprep without EDTA

The 260:280 ratio should  actually be as close to 1.8 as possible.  Lower is indicative of too much protein and higher can indicate RNA.  (Could always tell when the RNase was going bad when all the preps were 2.1 or higher.)  Anything (approximately) outside the 1.7 to 2.0 range and it becomes difficult to trust the 230nm reading used to calculate the concentration. 





On Saturday, July 28, 2012 7:09:44 AM UTC-7, Avery wrote:

Iirc as close to 2 as possible.

On Jul 28, 2012 5:38 AM, "Mega" <masterstorm123@gmail.com> wrote:

So, the values are:

For the 209.39 ng/uL plasmid  the 260:280 ratio is 1.46
176.51 ng/uL   -> 1.91
54.17 ng/uL  -> 1.74



I think 1.91 is the best? Should it be high or low? ;)





Am Sonntag, 27. Mai 2012 17:54:23 UTC+2 schrieb Mega:

Hello @all,

I was wondering if I could do a dirty miniprep without EDTA.

What I have: weak centrifuge, SLS (sodium lauryl sulfate), NaOH, vinegar, ethanol (I have access to both wodka and pure ethanol), water.


Can you do a mp with this limited resources?
What I want: Some ~60 to 70 % plasmids, some remaining proteins, RNA, mabe some fragments of chromosomal DNA.


I think with this percentage you can still do a transformation with E.Coli? Because Proteins will either be used or digested, RNA my do it's job inside the cell, chromosomal DNA has no origin of replication.

Only the plasmids will be able to replicate and thus you will get some transformants. These you select with ampicillin anyway. (Clearly, you won't get so much transformants, but who cares? )

Will DNAses inside the solution possibly destroy the plasmids??

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