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i still do my minipreps the old fashioned way, just add a chloroform extraction and PEG pptn step to get sequence quality DNA. It really doesn't take much longer than the kits, as you can do other things during the precipitation steps.
Also note that buffer p2 on the openwetware page should be made fresh each time as the NaOH gets neutralized by CO2 absorption from the air.
I also used to work in a lab that made their own (Promega) wizard miniprep columns using cheap filters and diatomaceous earth. I can't remember what sort of preparation was required though.
#139568 QIAGEN Spin Columns and Minipreps - Alternatives?!
Posted
on 16 August 2012 - 01:22 PM
#139505 Non-hazardous substitute for ethidium bromide?
Posted
on 15 August 2012 - 04:44 PM
Basically anything that binds to DNA has the potential to be hazardous, so it is unlikely that you will find something that is completely safe. There are a range of dyes that work with blue light illumination which cuts down on the UV exposure, and has the added bonus of not damaging the DNA when trying to see your bands prior to gel extraction.
#139464 QIAGEN Spin Columns and Minipreps - Alternatives?!
Posted
on 15 August 2012 - 05:45 AM
Epoch labs also sells inexpensive columns, and instructions on making the reagents.
See also: http://openwetware.o.../Qiagen_Buffers
See also: http://openwetware.o.../Qiagen_Buffers
#139364 Textbook Canon: Expressing Any Genes in Bacteria
Posted
on 13 August 2012 - 02:09 AM
Interesting.
*looks left and right* I think I'll have another go at it.
If all DNA/genes were destined for proteins, then there wouldn't be any tRNAs and rRNAs left. Or RNA I & II, and miRNAs for regulation [in C. elegans].
That should account for the difference in the number of genes and proteins.
I sure hope this is the 'simpler' answer
*looks left and right* I think I'll have another go at it.
If all DNA/genes were destined for proteins, then there wouldn't be any tRNAs and rRNAs left. Or RNA I & II, and miRNAs for regulation [in C. elegans].
That should account for the difference in the number of genes and proteins.
I sure hope this is the 'simpler' answer
#139327 question on symbols used in microbiology
Posted
on 12 August 2012 - 08:28 AM
I wrote a good portion of that page, and if you think things are unclear after reading it, you should have seen things before it existed. Genotypes are simply a mess. In the next few years we will mostly give up on them and start looking at complete sequence, which will tell what is really happening. Meanwhile, attempting to make more sense of the mess is, and will remain, very challenging. Figure out the properties you care about, and then test them on the strain you have in your hands.
