Question

What is ONE technique to determine the SEQUENCE of DNA/RNA AND proteins?

Answer 1

The most widely accepted method of DNA/RNA and protein sequencing method is the Sanger Sequencing method. Sanger sequencing begins with the denaturation of the double-stranded DNA. The single-stranded DNA is then annealed to oligonucleotide primers and elongated using a mixture of deoxynucleotide triphosphates (dNTPs), which provide the needed arginine (A), cytosine (C), tyrosine (T), and guanine (G) nucleotides to build the new double-stranded structure.

Protein Sequencing :

Edmans Degradation method is one of the most important reactions for protein sequencing. In this process, the ordered amino acid composition of proteins is discovered. Following steps are followed:
1. Disulfide bridges in the protein with a reducing agent such as 2- mercaptoethanol are broken down.
2. Then the individual chains of the protein complex are separated and purified.
3. Then the amino acid composition of each chain is determined.
4. The terminal amino acid of each chain is determined and then each chain is broken down into 50 long amino acids.
5. Then the fragments are purified.
6. The sequence of each fragment is determined and then the sequence of an overall protein is constructed.