Question

You have a sample that includes equimolar amounts of protein, dsDNA, ssDNA and typical eukaryotic mRNA. You want to separate the 4 components of the mixture in 3-4 steps, without destroying any of the molecules of your sample. At the end of your separation you must have one and only chemical species in each tube (one tube with dsDNA, one with ssRNA, etc.)

Briefly describe the sequence of steps that you would take using a diagram. To get full credit you need to:

• number the steps,
• sketch what you are doing,
• clearly and concisely explain what you are doing in words.

Answer 1

All four samples have different molecular weight and charge, so considering this we will do Native PAGE or 2-D PAGE,

1. Iso- electric focussing separate molecule based on the PI or charge and SDS-PAGE separate molecules on basis of Mass in 2-D PAGE.

2. After getting results, extraction of band got through PAGE and checking there purity with Nanodrop.

Mass and charge of protein, Ds DNA, ss DNA, and mRNA are different so we can separate them easily.