Hello everyone,
I am doing an electrophoretic mobility study on plasmid DNA (pBR322) treated with cisplatin.
The paper I am following says I have to dilute my DNA in a 1mM sodium cacodylate and 20mM NaCl buffer.
I understand the purpose of the NaCl, but not the sodium cacodylate. The literature I have so far come across tells me that this compound is often used for the preparation of microscopy samples, as it prevents the mitochondria and other organelles from rupturing. However, I was unable to find out what the purpose of this compound could be in an experiment involving free DNA (not cells).
They also use xylene (4mL!) to quench the reaction after incubation (I presume the reaction of cisplatin with DNA), and I am not sure why they do that either. Thought I would mention it, maybe it has something to do with the sodium cacodylate...
I would be very grateful if someone could point me in the right direction.
Thanks in advance!
Daniel D.
Sodium cacodylate is a common biological buffer; it has a good buffering capacity in the range of pH 5.0–7.4, so its role in the DNA isolation solution is to maintain pH. Cacodylate is often used instead of the phosphate buffer in the applications where it is undesirable to have phosphates introduced into the sample.
Sodium cacodylate is a common biological buffer; it has a good buffering capacity in the range of pH 5.0–7.4, so its role in the DNA isolation solution is to maintain pH. Cacodylate is often used instead of the phosphate buffer in the applications where it is undesirable to have phosphates introduced into the sample.
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