The Sanger Coulson method includes DNA denaturation, PCR with ddNTPs, further denaturation, and polyacrylamide gel electrophoresis to determine the DNA sequence.
The Sanger Coulson method involves DNA denaturation, PCR amplification with ddNTPs in four tubes, subsequent denaturation, and separation on polyacrylamide gel to read the sequence from bottom to top.
| Front | Sanger Coulson method steps |
|---|---|
| Back | 1) DNA is denatured using heat and a single strand is obtained that is tagged with a known sequence at the 3' end. 2) The tagged DNA strand, Taq polymerase, Primers and Nucleotides are added in four tubes, each containing a different ddNTP, and placed in PCR machine. 3) The product DNA is denatured again and each tube is run into a different lane of Polyacrylamide gel 4) The sequence is read from bottom to top and then its complement is deduced. |
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