Question

I'm digesting a plasmid for assembly. In my experience, I always have low yields when gel extracting and purifying. Since I am only making one cut in my vector, could I just purify with a column?

Answer 1

Yes, you can purify the plasmid DNA with a column if you are only making one cut in your vector. Column purification using silica-based anion-exchange chromatography is a faster and more efficient method. However, it's important to follow the plasmid prep protocol accurately to achieve optimal yield and purity.

Step-by-step explanation:

When purifying plasmid DNA, gel extraction is a common method used to isolate the desired DNA fragment. However, if you are only making one cut in your vector, it is possible to purify the DNA using a column instead.

Column purification, specifically using silica-based anion-exchange chromatography columns, takes advantage of the negative charge of DNA to separate the plasmid DNA from the host's genomic DNA. This method provides a faster and more efficient way to purify the plasmid DNA.

It's important to note that the yield and purity of the DNA can depend on the specific plasmid prep protocol you are using. Following the protocol accurately, especially the initial steps, can significantly impact the outcome.