Cloning and Mutation Service
01. Cloning of DNA Fragment into Bacterial and or Mammalian Vectors from cDNA or Genomic DNA Cat # BB-SC10
Template DNA of any desired length will be cloned into a vector of choice. Bio Bharati can provide generic pUC / pET vectors or Bio Bharati developed specialized vectors to generate the desired clone as per requirement. Template DNA will be generated from customer specified sources or can be engineered by fusion of DNA fragments according to the need of the customer. Customer may also provide the specific vectors of their choice. The clone will be confirmed by Sequencing.
Note: Cell lines if required for cloning purpose need to be provided by customer.
02. Mutagenesis to Create Mutants Cat # BB-SM20
PCR-Based method with Pfu Polymerase will be used for the generation of mutants. Insertions / Deletions / Substitution of single/multiple bases can be generated as per need. Customer will provide the template. Bio Bharati will design and synthesize the required primers. Mutations will be confirmed by Sequencing.
Protein/ Antibody coupling to Agarose Beads:
01. Immobilization of protein on agarose beads: Cat # BB-SIA45
Protein of interest (3 to 8 mg) will be coupled to 1 to 5 mL of cross-linked agarose bead (pack volume) depending on customer need.
02. B. Immobilization of antibody on Agarose Beads: Cat # BB-SIA45
Antibody of interest (3 to 8 mg) will be coupled to 1 to 5 mL of cross linked agarose bead (pack volume) depending on customer need.
Protein Expression, Purification and Optimization
Expression Optimization of Recombinant protein in E.coli system Cat # BB-SPE30,
Protein Purification from Recombinant source Cat # BB-SPP40 & Protein Purification from natural sources Cat # BB-SPP50, BB-SPP60
We express and purify protein of interest from E.coli following standard optimization procedures. If the expression is optimized by the customer, we will follow the same procedure to scale up protein expression. We also purify protein of interest from natural sources (plant or animal origin). Amount of protein delivered will depend on level of expression and solubility.
Antibody generation and conjugation
Antibody generation and conjugation Cat # BB-SAP50
Two rabbits will be injected with protein and peptide antigen. We will provide about 20 mL of antiserum or ≥2 µg affinity purified antibody as per customer need. Customer should provide soluble protein antigen (~5 mg for antiserum and 10 mg for true affinity purified antibody) or peptide antigen with one extra cysteine residue at the N or C terminal end for KLH conjugation (~5 µg for antiserum and 10 µg for affinity purified antibody). We will procure the protein or peptide with extra charges.