Description:The PCR Cloning Kit is a simple and highly efficient method to insert any gene or DNA fragment into a vector, without time-consuming ligation steps
No ligation steps required
Multiple PCR inserts can be added in a single step
Efficiencies of > 90% are typical.
Go from PCR product to transforming cells in under an hour!
Suitable for fixing or adding point mutations, insertions or deletions.
Test Principle:A primer set homologous to the gene of interest and the vector is used to amplify a PCR product. This purified PCR product is incubated with restriction enzyme-digested vector and Reagent A of the PCR Cloning BioAssay™ Kit. Following a brief incubation with Reagent B, the annealed vector is ready for transformation.
Primer Design:Proper primer design is critical for successful PCR and transformation steps. Primers for the gene of interest should contain the restriction site as well as at least 10 nucleotides of sequence homologous to the vector sequence. Using two different restriction enzymes will allow directional cloning of the insert. Note: Once purified, the digested vector can be used with multiple inserts if the primers for insert amplification are designed correctly.
Kit Components:137318A: Reagent A, 1x1vial137318B: Reagent B, 1x1vial137318C: Buffer A, 1x1vial137318D: BSA, 10X (0.1% w/v), 1x1vial137318E: Distilled Water (dH2O), 1x1vial
Storage and Stability:Store all components at -20°C. Stable for 6 months after receipt. Aliquot into working volumes after first thawing of components to avoid multiple freeze-thaw cycles. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
