What is the purpose of resuspending your pellet in low salt

What is the purpose of resuspending your pellet in low salt buffer C of 0.025M KCL after 1st centrifugation? Was Taq DNA polymersase in the pellet or supernatant?

Solution

1.Resuspending pellet in low salt buffer after 1^st centrifugation-

First we need to centrifuge bacterial culture for removal of undesirable cell debris or biomolecule like protein, carbohydrate, lipid, than we resuspending this pellet in low salt buffer for important reason, when we add resuspend pellet in low salt than the DNA of cell make ionic bond with salt and remove the water molecule surrounding DNA and the DNA have phosphorus –ve charge group (-PO4)that will make ionic bond with salt so after that DNA interacting with salt and resulting in DNA-salt settled down on bottom, so that make all the DNA in bottom of tube with salt, so there is a maximum chance to get our desirable DNA in pellet form, for good PCR amplification result, that’s way resuspending in low salt is important for our experiment.

2. Was Taq DNA polymersase in the pellet or supernatant ?

Taq DNA polymerase present in the PCR reaction pellet not in supernatant.

And PCR reaction pellet also contain Mg 2+, four dNTP’s, buffer, and you make sure the reaction pellet is at the bottom. And we know the importance of this reaction pellet for very sensitive PCR amplification,Taq DNA polymerase role is extension of DNA strand both side and Mg2+ is cofactor of polymerase is crucial for polymerase function, and all four dNTP’s required for DNA amplification