Question

1) A higher concentration of agarose (and hence smaller pore size in the gel) will allow you to separate what more efficiently? please list examples with explanation.

2) What could be a problem when trying to clone a 3.5kb fragment into a 3.5kb vector? Examples and explanations please.

Thank you.

Answer 1

Ans. Agarose gel is used for the separation of nucleic acids and proteins. The separation of different sizes molecules is depends on the concentration of agarose in gel. Higher the concentration of agarose, lesser will be the size of pores of gel. Smaller the size of pores allow separation more efficiently. For example, larger nucleic acid molecules (such as genomic DNA) is separated by 0.8% gel, while smaller DNAs (such as PCR products) are separated by 2% gel.