The laboratory technique involving the extraction of DNA from a cell in a pure form.
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2answers
516 views
How do I prepare and clone from E. coli DNA?
I'm looking for a protocol to get genomic DNA from an E. coli sample so that I can clone a small portion of it using PCR into a plasmid. (< 500 bp in this case).
It seems OWW (Open Wet Ware) ...
4
votes
2answers
483 views
Why is proteinase K digestion performed at 50 °C?
Many DNA isolation protocols use a Proteinase K digestion to remove proteins. This is often performed at 50 °C. Why is this?
8
votes
1answer
464 views
How to DIY preserve pet DNA today so that it can be used in 20 years
I know that there are companies that offer a kit+storage of your pet DNA for around $2k.
My question is if there is any other option for doing this yourself with a thought in mind that in 20+ years ...
9
votes
2answers
214 views
What is causing my problem with very low yields when isolating a 42kb yeast plasmid?
I have to isolate a large plasmid from yeast and transform it in E. coli. After transformation, I often get no colonies. One reason for that is the yeast mini prep hasn't worked or the DNA ...
8
votes
1answer
3k views
Why do we add salt when precipitating DNA?
All the DNA extraction protocols I have seen involve adding salts to the extraction buffer. What is the purpose of the salts? What happens if they aren't included?
4
votes
4answers
291 views
What is a good miniprep protocol for the class room?
I'm trying to find a good protocol for plasmid minipreps and I'm looking at 3 preps I've found:
Using phenol/chloroform
extract with phenol:chloroform:isoamylalcohol,
isopropanol precipitation, ...
