Catalog #24101 - Cloning and sequencing of mouse hybridoma IgG variable regions
Title:
Cloning and sequencing of mouse hybridoma IgG variable regions
Description:
Cloning and sequencing of mouse hybridoma IgG variable regions
Timeline:
1 Week
Deliverables:
1. Study report containing sequence analysis. 2. Amplified PCR fragments of the variable heavy and light chain sequences can be provided for an additional $100 plus shipping/handling.
Material Required:
A. Greater than 100,000 cells in frozen vial(s) and shipped on dry ice
OR B. Isolated RNA samples dried on RNA isolation columns (before elution) and shipped on dry ice.
The recommended protocol for sample preparation:
Treat your hybridoma sample with RNAlater stabilization solution and then frozen for shipping on dry ice.
(https://www.thermofisher.com/order/catalog/product/AM7020)
Scope:
Cloning and sequencing antibody variable regions:
1. Extract and purify total RNA from hybridoma cells or clonal B cell.
2. Synthesize cDNA using optimized primer set.
3. Amplify the DNA fragments corresponding to the VL and VH using nested PCR.
4. Sequence the amplified DNA fragments. Amplified PCR fragments of the variable heavy and light chain sequences can be provided for an additional $100 plus shipping/handling.
Deliverable:
1. Study report containing sequence analysis. 2. Amplified PCR fragments of the variable heavy and light chain sequences can be provided for an additional $100 plus shipping/handling.
Material Required:
For cloning and sequencing antibody V regions: A. Greater than 100,000 cells in frozen vial(s) and shipped on dry ice OR B. Isolated RNA samples dried on RNA isolation columns (before elution) and shipped on dry ice.
Timeline:
1 week
Price:
$1,500