This is a product manual that introduces the use of pRSET A, B, and C plasmids, including cloning, expression, and purification.
This manual provides instructions for the Invitrogen V351-20 kit, which includes pRSET A, B, and C plasmids designed for high-level protein expression and purification in E. coli. The vectors utilize a T7 promoter and an N-terminal fusion peptide containing a polyhistidine tag and an Xpress epitope.
| Plasmid Concentration | 0.5 μg/μl |
|---|---|
| Plasmid Volume | 40 μl each |
| Antibiotic Selection | 50–100 μg/ml ampicillin |
| Buffer pH | 8.0 |
The manual recommends using a recA, end A strain such as TOP10F' for cloning and transformation.
Upon receipt, the plasmids should be stored at –20°C.
The polyhistidine tag functions as a metal binding domain to allow simple purification of recombinant proteins by Immobilized Metal Affinity Chromatography.