ALU POLYMERASE CHAIN REACTION LAB
Purpose: The purpose of this lab is to prepare a PCR, polymerase chain reaction, for an amplification of an Alu insert after isolating DNA from cheek cells.
Materials: Pipets(2-20; 20-200; 100-1,000 micro-liters), Pipet tips, Salt water, Micro centrifuge, Wast containers, Micro centrifuge tubes, PCR tubes, Agarose, TAE buffer(1x), Gel tanks, Gel molds, Loading dye, Tube racks, Primer mix, Master mix(DNA polymerase, reaction buffers), Distilled water, DNA(positive control), Sharpie, Chelex, Heat block, Thermal cycler
Materials: Pipets(2-20; 20-200; 100-1,000 micro-liters), Pipet tips, Salt water, Micro centrifuge, Wast containers, Micro centrifuge tubes, PCR tubes, Agarose, TAE buffer(1x), Gel tanks, Gel molds, Loading dye, Tube racks, Primer mix, Master mix(DNA polymerase, reaction buffers), Distilled water, DNA(positive control), Sharpie, Chelex, Heat block, Thermal cycler
Procedure- !st swish salt water around 30 sec, 2nd Microfuge, add saline, rack cells, drain out some saline, re-suspend cells, add chelex, re-suspend, heat for 10min, open to release pressure, get cells and put into new tube and transfer, refrigerate, add master mix to new tube, then add primer, add dna to new tube, put into heater, -95Cfor 2 minutes then 30 cylces of 94C for 30 sec, 60C for 30 sec, 72C for 2 minutes, seperate cycle72C for 10 min, 4C infinity hold