Improve: PCR product purification

#PCR product purification is as important and tricky as adding egg whites into your #WhiskeySour. Nevertheless all the downstream applications of the PCR product depend largely on the quality of the purified #DNA. Linear #acrylamide can serve as a very good enhancer here.

Protocol

1. The Sample DNA needs to be in a salt solution, or you may add 3M #SodiumAcetate pH 5.2 to it 1/10th the sample volume.

2. Add 10-20ug/mL of linear #Acrylamide to the sample.

3. Add one volume of #Isopropanol (preferably) or two and half volume of #Ethanol.

4. Mix and keep in -20C freezer for at least 15 mins (overnight would be best though!).

5. Centrifuge at ~12000g at 4C for 10 minutes.

6. Discard the supernatant and wash the visible pellet with 70% #Ethanol.

7. Repeat centrifugation, dry the pellet and dissolve in suitable buffer or water.

Merits

1. Linear #acrylamide can improve yield of >20bp PCR products, excluding most #oligos and #dNTPs.

2. Being an artificial chemical it does not have chances of being contaminated with other #NucleicAcids or #nucleases as is the case with other popular choices like #YeastRNA, #tRNA and #Glycerol.

3. #Acrylamide does not interfere with the A260/280 ratio, although #Glycerol might.

4. #Acrylamide does not interfere with DNA during phosphorylation with polynucleotide kinase like #tRNA or with DNA-protein interaction experiments like #Glycerol.

More to chew:

# https://patents.google.com/patent/US5422241A/en