Pre-Amplification Protocol for PicoPLEX WGA Kit

Protocol

1. Combine Extraction Cocktail components and mix well:

   EXTRACTION COCKTAIL	VOLUME PER 5 SAMPLES
   Extraction Enzyme Dilution Buffer	24 μl
   Cell Extraction Enzyme	1 μl
   Total volume	25 μl

2. Add 5 μl of freshly-prepared Extraction Cocktail to each 5 μl cell sample or control DNA sample prepared by the sample preparation methods.
   
   
3. Incubate sample in a thermal cycler as follows:
   

   CYCLES	TEMP	TIME
   1	75°C	10 minutes
   	95°C	4 minutes
   	12°C	Hold

4. Combine Pre-Amp Cocktail components and mix well:

   PRE-AMP COCKTAIL	VOLUME PER 5 SAMPLES
   Pre-Amp Reaction Mix	24 μl
   Pre-Amp Enzyme	1 μl
   Total volume	25 μl

5. Add 5 μl of Pre-Amp Cocktail to each cell or control sample.
   
   
6. Incubate sample according to the thermal cycler program below:
   

   CYCLES	TEMP	TIME
   1	95°C	2 minutes
   12	95°C	15 seconds
   	15°C	50 seconds
   	25°C	40 seconds
   	35°C	30 seconds
   	65°C	40 seconds
   	75°C	40 seconds
   1	4°C	Hold

7. Briefly centrifuge sample and place Pre-Amp incubation product on ice.