Objectives:
• To perform PCR amplification of specific target sequence from template DNA.
• To analyzed the amplified product by Agarose gel electrophoresis.
Principle:
PCR-polymerase chain reaction is an in vitro method of enzymatic synthesis of specific DNA sequence, developed by Kary Mullis in 1983. It is a very simple technique for characterizing, analyzing & synthesizing any specific DNA or RNA from any source.
Materials Quantity Storage Condition
Taq DNA polymerase 5µl -20°C
10X Taq Buffer 50 µl -20°C
Template DNA 5µl -20°C
Forward Primer 5µl -20°C
Reverse Primer 5µl -20°C
Nuclease free water 200 µl -20°C
1kb DNA ladder 25 µl -20°C
dNTP mix 10 µl -20°C
6X DNA Loading Dye 25 µl -20°C
Agarose 1 gm RT
50X TAE 7.5 ml 4°C
PCR Tubes 5 Nos. RT
Ethidium bromide (EtBr) 25ul RT