The first step in this process is identification of the regions of the DNA that are going to be used in the synthesis. This is done by use of bioinformatics. When these regions are synthesized they form antigenic fragments. The region that binds to the antigen on an antibody is known as the fragment. It has a single constant and variable domain of the chain (Talpaz, 2003).
The second step is the design of polymerase chain reaction primers. The primers are used to amplify these regions. Primers are used as the starting point to synthesis of DNA. The polymerase chain reaction is used to magnify DNA strands through several orders of magnitude. This leads to the replication of the DNA. This process consists of a subsequent repetition of 20-40 temperature changes. These steps known cycles consist of 2-3 discrete temperature steps. This process happens in three stages:
- Exponential amplification: This involves doubling the product in every cycle.
- Leveling off stage: This involves slowing down of the reaction until the DNA polymerase loses activity.
- Plateau: There is no more formation of products as the entire reagent and enzyme for the reaction is used up (McFarland, 2009).
The third step involves cloning of the DNA using different expression systems. The regions of the DNA that were amplified are first purified using a gel in preparation for the cloning stage. They are then isolated and after that screened for expression performance (Perrone, 2005).
Sequencing and Alignment Analysis is the fourth step in this process. The DNA is arranged in such a way that the regions will be similar in function and structure.
The fifth step is to identify the clones that are relevant for the synthesis. This is can be done in two ways depending on the expression yields required (Kitcher, 2007).
The two are Higher-plex techniques and Low-to-mid-plex techniques.
After that the conditions and additives are optimised so as to improve on the yield of the clones.( Othman& Hart, 1988).
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