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Preparatory Steps

 laminar flow hoodAll work from this point on will be done inside a laminar flow hood. So the first step here is to remove everything from within the hood and thoroughly spray all surfaces with 70% alcohol and wipe down.

ALL reagents, master mix coctail, DNA, PCR products, and 0.2 mL reaction tubes should be stored on ice at all times or in a benchtop PCR cryo-pak.

Place ice buckets and all items listed above in hood and turn on UV light for at least 10 minutes (even just walking through the lab with any of these materials exposed to the air can introduce surface contamination—hence the UV exposure).

Preparation of Master mix cocktail

Loading microfuge tubes for PCR reaction