RT-PCR

RT-PCR is a pretty standard procedure, and now its use is extremely broad. At its most basic application, PCR can be extended to small amount of template DNA (or RNA) in large quantities within a few hours. This is done by mixing the DNA on both sides of the DNA primers (reverse), Taq polymerase (Thermus aquaticus type, polymerase thermophile that is able to withstand very high temperatures), free nucleotides (dNTPs on DNA, RNA, NTPs), and buffer. The temperature is then denatured, and alternating hot and cold reanneal DNA polymerase, the addition of new complementary strands each time. In addition to the main use of PCR, which is specially designed primers will ligate to two different pieces of DNA together, or add the restriction site, and many other creative applications. It is clear that PCR is a procedure that is an integral addition to molecular biology tools and techniques are constantly improved over the years.