- Step 1: Obtain all necessary components to make master mix (buffer, polymerase, nucleotide-free water, reverse and forward primer pair, dNTP mix) from block in freezer.
- Substep 1: Collect ice from down the hall in a clean PCR-only bucket. Put all ingredients into this bucket immediately, except the frozen buffer: warm this up to 37 degrees.
- Substep 2: Label the 8 thing.
- Substep 3: Make sure the correct PCR program is running.
- Preparation must be done inside the sterile PCR hood
- Calculate all necessary amounts of ingredients
- Make sure to add one more unit than needed as wiggle room
- Step 2: Make master mix, changing pipette tip for each ingredient.
- Step 3: Take master mix out of hood and transfer ice into non-clean (green) bucket.
- Step 4: Make sure MM is well mixed using the pipette. Transfer into the 8 thing.
- Step 5: Transfer extracted mouse DNA into the 8 thing, changing the pipette tip each time.
- Step 6: Place into thermocycler.
- Step 7: Clean work area and place all ingredients back into their correct places. Do not leave polymerase out on the counter, it will denature and it's RIDICULOUSLY EXPENSIVE TO GET MORE! LIKE $1000 FOR A TINY TUBE
aug 8 2015 ∞
aug 29 2015 +
