Microsphere Polymerase Chain Reaction to Amplify Single-Stranded DNA

Obtain approximately 25 microspheres through microscopic counting using a light microscope with 40X magnification. Following this, combine all reagents as described in the text protocol. Place the samples into a thermocycler, and start the asymmetric PCR under the appropriate conditions.

The collection of supernatant after a symmetric PCR is one of the most critical steps because it is a major process for generating single-strand DNA with microspheres.

Following amplification, add eight microliters of 6X loading buffer to each sample. Load 15 microliters of each sample into 2% agarose gel. Then, perform electrophoresis at 100 volts for 35 minutes in 1X TAE buffer.