Subcloning into Expression Vector
- Amplification/isolation of the gene of interest from a customer-supplied vector and subcloning into a suitable expression vector.
- Verification of the subcloned gene by restriction digest and sequencing.
- Preparation of the recombinant DNA construct.
- Transformation of recombinant constructs into a suitable Picha strain.
- Small scale induction to over-express target protein.
- Test for recombinant protein by SDS-PAGE and western blot (customer must provide appropriate antibodies)
Estimated time: 3-4 weeks
Generation and Identification of Expression Positive Clones
- Large scale induction of clonal populations and 1L culture to over-express target protein.
- Ni-NTA purification of protein
- SDS-PAGE and WB confirmation of purified protein
- Delivery of purified protein in PBS or customer-specified buffer
Estimated time: 2 weeks