TACGen provides sequencing service for monoclonal antibody hybridoma cells.
Starting Materials:
A pellet of hybridoma cells (1x107 cells) expressing your antibody
An EBV transformed B-cell line
Technical Steps:
RNA Preparation:
RNA will be extracted and purified from the hybridoma cell pellets.
cDNA Synthesis:
Reverse transcrition will be perfomred to synthesize cDNA. Specific murine and human constant domain primers can be used to determine the isotype of the antibody.
PCR or 5'RACE: PCR products will include the regions of the signal peptide, variable domains and constant domains up to the reverse primer.
Cloning:
The PCR products will be gel purified and clone into a sequencing vector for sequencing.
Sequencing:
A minimum 10 clones will be sequenced for each chain.
Report:
A detailed report will be produced including the sequence alignments of the heavy and light chains.
