Amplification Protocol for PicoPLEX WGA Kit
- Combine the following Amplification Cocktail components and mix well:
AMPLIFICATION COCKTAIL VOLUME PER 5 SAMPLES Amplification Reaction Mix 125 μl Amplification Enzyme 4 μl Nuclease-Free Water 171 μl Total volume 300 μl
- Mix 60 μl of the freshly prepared Amplification Cocktail with the 15 μl pre-amp incubation product and mix gently by pipet.
- Amplify sample according to the thermal cycler program below:
2 minutes 14 95°C 15 seconds 65°C 1 minute 75°C 1 minute
Note: 14 cycles are recommended based on testing performed with flow-sorted cultured cells. Some cell types may require up to 16 cycles to obtain maximal yields.
- Immediately store the amplified product at -20°C or purify.
Note: Many applications require purifying and quantifying WGA products before use. Products amplified using this kit can be purified with spin columns or filter plates.
- Quantitate purified amplification products by UV absorbance (1 OD260 = 50 μg/ml). Store the purified amplification product at -20°C.
Note: PicoGreen™ or other double-stranded specific methods for DNA measurement will not give reliable product concentrations. The amplified material will be a mixture of predominantly double-stranded material with some single-stranded material present.