IgG, Library Primer Set, Mouse, BioGenomics™

Kits and Assays / Kits-Cloning, BioGenomics™

-20°C

Each primer is supplied as a liquid in 10mM Tris-HCl pH 8, 1mM sodium EDTA.

Many monoclonal antibodies of mouse origin are valuable diagnostic agents. Their production by classical hybridoma techniques is frequently limited by the instability of cell lines, low antibody yields and the limitations of immunizing mice with toxic antigens. A promising alternative to the hybridoma technology is the production of recombinant antibodies. Pioneering work of the last decade showed that it is possible to amplify rearranged immunoglobulin genes from B-lymphocytes, to insert them into different vectors, and to express them in bacteria, yeast, insect, mammalian or plant cells. Moreover, the randomized combination of cloned heavy and light chain immunoglobulin gene fragments allowed the construction of mouse antibody libraries. These libraries enable the isolation of specific antibodies against particular antigens by phage display techniques. One prerequisite for generating highly diversified mouse antibody libraries, however, is the development of PCR primers capable of amplifying all rearranged immunoglobulin genes. In immunoglobulin repertoire library cloning, the homology between a particular primer sequence and its target template, as well as the diversity of a primer pool are the two most important parameters which determine the cloning efficiency and the size of a resulting repertoire library. This screening strategy allows the amplification of rearranged mouse immunoglobulin genes of individual B cell clones as well as of larger B cell populations for the construction of mouse scFv-antibody libraries.

Kit Components:
Primer Set 1 for PCR Amplification
I1904-10A1: Heavy Chain Variable Primer (Set 1) 11x20ul
I1904-10A2: Heavy Chain Constant Primer (Set 1) 1x220ul
I1904-10A3: Light Chain Variable (kappa) Primer (Set 1) 10x20ul
I1904-10A4: Light Chain Constant (kappa) Primer (Set 1) 1x220ul
I1904-10A5: Light Chain Variable (lambda) Primer (Set 1) 1x20ul
I1904-10A6: Light Chain Constant (lambda) Primer (Set 1) 1x20ul
Primer Set 2 for Cloning
I1904-10B7: Heavy Chain Variable Primer (Set 2)11x20ul
I1904-10B8: Heavy Chain Constant Primer (Set 2) 1x220ul
I1904-10B9: Light Chain Variable (kappa) Primer (Set 2) 10x20ul
I1904-10B10: Light Chain Constant (kappa) Primer (Set 2) 1x20ul
I1904-10B11: Light Chain Constant (lambda) Primer (Set 2) 1x220ul
I1904-10B12: Light Chain Constant (lambda) Primer (Set 2) 1x20ul

PCR amplification (Set 1) and cloning (Set 2) of mouse IgG heavy and light chain variable domain coding regions.

Applications:
Generation of large repertoires of rearranged immunoglobulin variable domain coding regions for the construction of mouse IgG scFv-antibody libraries. Amplification of immunoglobulin variable gene fragments from single B cell clones.

Recommended Dilutions:
Working dilution 10pmol/ul (Ready-to-use for common PCR).
Dilution buffer H2O.
Category PCR oligonucleotide primer.

Storage and Stability:
Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for 6 months after receipt.