Catalog #24102 - Cloning and sequencing of rat hybridoma IgG variable regions

Title:
Cloning and sequencing of rat hybridoma IgG variable regions

Description:

Cloning and sequencing of rat hybridoma IgG variable regions


Timeline: 2 Weeks


Deliverables: Study report containing sequence analysis. 

Note: Amplified PCR fragments of the variable heavy and light chain sequences can be provided for an additional $100 plus shipping/handling.


Material Requirement:

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.


suggest client treat their sample with RNAlater stabilization solution (https://www.thermofisher.com/order/catalog/product/AM7020), and then frozen and send out without concern for cell rupture and release of RNases since the RNases have already been inactivated



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:
1-2 weeks

Price:
$2,400