Explain in general terms what is happening in each of the fo

Explain in general terms what is happening in each of the following steps of the miniprep DNA purification. It may be necessary to look online or in the Qiagen manual posted on Canvas to understand what happens in each step based on solution compositions.

1. Cell Resuspension

2. P2 buffer addition

3. N3 buffer addition

4. PB column wash

5. PE column wash

Solution

1. Cell Resuspension-This is a process which basically involves breaking up the cell pellet or the DNA pellet.Moreover It involves the process to get the various kinds of cells back into solution.It further involves vortexing the sample of the given DNA.

2.P2 Buffer Addition:

It is defined as a process in the formation of Clumps that takes place on the addition of Buffer P2 in a bacterial lysate.This bacterian lysate usually contains LyseBlue reagent which is the biggest indicator of the fact that there is ia poor resuspension of the bacterial cell pellet in Buffer P1. This causes handling error which usually leads to inefficient cell lysis.It also cuses and genomic DNA and cell debris.

3.N3 buffer addition:In this process we will add the Buffer N3.This will help to neutralize the lysis reagent,Then we will mix the mixture immediately by inverting the tube six times. we will not vortex here and finally we will Centrifuge the mixture which we have with us for 10 minutes.

4.PB column wash:In this process we will add the Buffer PB to the spin cartridge.This is followed by Centrifugation at maximum speed for 1 minute. The main purpose is to discard the flow-through generated here.

5.PE column wash:In this process we will add the PE to the given cartridge. This is followed by spinning for 1 minute and finally discarding the flow-through. The next step is to Spin again for 1 min removing the residual buffer.