AMPure PCR Purification Kit Sample Clauses

AMPure PCR Purification Kit. The PCR plate was centrifuged briefly using a plate centrifuge to ensure all PCR product is collected in the bottom of the well and also to reduce the risk of cross contamination. The seal was carefully removed from the PCR plate to avoid splashing. After gently shaking the AMPure XP magnetic particle solution container (kept at 4 oC but brought slowly to room temperature over a 20 minute period) to re-suspend any magnetic particles, using a multipipette 1.8 μl of the solution/1.0 μl of PCR product were added to the well. One negative PCR control was used, to help identify any potential contamination. We pipetted up and down gently x10 times to mix the beads and PCR product. The ensuing homogenous mixture was left to stand at room temperature for 5 minutes to enable DNA products (≥ 100 bp) to bind to the magnetic beads. The reaction plate was placed onto a 96 well magnetic plate for 5 minutes to separate the beads from the solution. Beads formed a ring around the well at the height of the magnet, the solution at the end appearing clear. With the reaction plate still on the magnetic plate the supernatant was pipetted and discarded, and a multipipette was used to add 100 μl of 70% ethanol solution to each well. The mix was incubated for 30 seconds and ethanol was aspirated and discarded. This process was repeated twice. Any remaining ethanol was aspirated and discarded by using a multipipette with 10 μl tips to avoid any PCR inhibition caused by the presence of ethanol. The plate was removed from the magnetic stand and was left uncovered to dry for 10 minutes avoiding any cracking of the bead ring, as this would significantly reduce the efficiency of the elution. Thirty μl of distilled water were added to each well with a multipipette. A mix was created by gently multipipetting up and down for 10 times. The reaction plate was put onto the magnetic platform for another 5 minutes to separate the beads from the solution.