How to conduct proper antibody sequencing library? Can anybody help me? Thanks a lot!
To conduct a proper antibody sequencing library, you need to avoid the following drawbacks, such that: inaccuracy of full sequence, PCR mutations, hybridoma cells impurities.
The accuracy of a full sequence is obtained if the individual sequences resulted from each chain are properly lined up.
PCR mutations occur when degenerate primer sets are used, during the development of cDNA.
In the structure of hybridoma, there exists the possibility of find mRNAs non-functional encoding antibodies, that can be developed by primer pairs, whose aim is to copy the targeted antibody.