Monoclonal Antibody Sequencing Service
Catalog No. |
Description |
VDA-100 | Variable Domains Sequencing Service -Total RNA extraction from the hybridoma cell pellet
-Reverse transcription
-PCR using degenerate primers for variable domains
-Cloning of the variable heavy chain sequence
-Cloning of the variable light chain sequence
-Final report (see example) | VDB-100 | Variable Domains & Leader Sequence Sequencing Service -Total RNA extraction from the hybridoma cell pellet
-Reverse transcription
-5RACE amplification of the heavy and light chain from antibody constant domains CH1 and CL
-Cloning of the variable heavy chain sequence
-Cloning of the variable light chain sequence
-Final report (see example) | VDC-100 | Variable & Constant Domains, and Leader Sequence Sequencing Service -Total RNA extraction from the hybridoma cell pellet
-Reverse transcription
-5RACE amplification of the heavy and light chain from antibody constant domains CH1 and CL
-Cloning of the variable heavy chain sequence
-Cloning of the variable light chain sequence
-Final report (see example) |
Descriptions:
MCLAB offers a fast and professional sequencing service for your valuable monoclonal antibodies. We can sequence from any antibody producing cell line with a service that can be tailored to your requirements for Quality Control, Patent Applications, Full Traceability and Drug Development.
MCLAB has extensive experience in antibody V-region determination from cDNA sequencing . Sequences are compiled and aligned based on bi-directional sequencing of multiple independent clones. We offer a rapid yet high quality and cost effective service.
Features: Personalized Ph.D. level project support for your inquiries: virtually no project limitations in terms of size and complexity The lowest price and the fastest monoclonal sequencing service on the market: as a global market leader in DNA sequencing services and consumables , we offer you fastest turnaround times and highest sequencing capacities on the market Reliability and productivity based on technological experience
Starting Materials - A pellet of snap frozen cells (1x107 cells) expressing your antibody - An EBV transformed B-cell line
Technical Summary
Stage 1: Total RNA extraction from the hybridoma cell pellet
Total RNA will be extracted and purified from the hybridoma cell pellet. The quality of the total RNA will be assayed on the Agilent Bioanalyzer 2100.
Stage 2 : Reverse transcription
Total RNA will be transcribed into cDNA using either an Oligo(dT) or a gene-specific anti-sense primer. Specific murine and human constant domain primers can be used to determine the isotype of the antibody.
Stage 3 : PCR or 5 RACE amplification of heavy and light chains
Degenerate VH and VL primers are used to amplify the variable domains from the cDNA.
For 5RACE, our proprietary adaptor is added to the 3 end of the cDNA. The heavy and light chains can now be amplified with our proprietary adaptor (sense primer) and a gene specific (CH/CL, reverse primer). PCR products will include the sequence of the signal peptide, variable domains and constant domains up to the reverse primer.
Stage 4 : Cloning into a standard sequencing vector
The PCR products will be gel purified to clone into a sequencing vector for sequencing.
Stage 5 : Sequencing analysis
As a standard, we will sequence a minimum of 12 independent clones for each chain.
Stage 6 : Final Report
A detailed report is produced on the work performed which includes the sequence alignments of the heavy and light chains and is e-mailed to the client.
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