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Real Time PCR

The main purpose of Real Time PCR is to amplify the number of copies of a specific region of DNA, in order to produce enough DNA to be tested as needed. This technique is useful as it can be used in many different circumstances such as: to identify with a very high-probability, disease-causing viruses and/or bacteria, a deceased person, or a criminal suspect.

Advantages of Real Time PCR

Efficient

- Incredible amplification. One molecule can produce large amounts of product.

Specific

- can amplify desired sequence only.

Problems in Real Time PCR

Contamination

- one contaminating molecule of DNA can produce non-specific bands. Solution-: Use sterile technique & keep prep and PCR amplification in separate rooms.

Miscorporation - Taq Polymerase has no proof reading ability. Solution-: sequence more than one clone or use Vent Polymerase, which has proof-reading abilities.