PCR clean up using 3M sodium acetate and chilled absolute ethanol
For each 20 ul of PCR product, prepare the following mixture in 1.5 ml Tube.2 ul
For each 20 ul of PCR product, prepare the following mixture in 1.5 ml Tube.
- 2 ul of 3M sodium acetate (NaOAc) pH 4.5
- 40 ul chilled absolute ethanol.
Transfer the PCR product (20ul) into the tube containing the mixture. Vortex the tube and then store it at -20 for 30-50 min. Spin for 30 min at maximum speed 14.000 rpm. Rremove the supernatant carefully and don''t disturb the pellet. ( Do not touch the bottom of the tube) Wash the pellet with 70% ethanol. And spin the tube at maximum speed for 5 min. Remove the supernatant and then dry the pellet using vacuum centrifuge. (do not apply heat) Resuspend the pellet in 30 ul in sterile water.