SensiFAST™ Probe No-ROX Kit, from Bioline for End-Point PCR – Bioline
Bioline’s SensiFAST™ Probe No-ROX Kit, from Bioline utilizes the enzyme included in the components, to amplify DNA, and similar templates. Included PCR reagents and buffers help support regular PCR in addition to singleplex PCR, allowing complete control over amplification specificity (i.e., specific or non-specific amplification) and complexity (single or multiple target amplification).
Polymerase and Protocol Specifications
The polymerase is supplied at the specified concentration and is activated by conditions stated in the protocol during denaturing. Following annealing, nucleic acid samples of the suggested weight or volume are elongated at the optimum temperature. Bioline recommends a suitable PCR reaction volume and amplicons of appropriate size for optimal polymerase performance. Utilizing thermal cycling and regular PCR to replicate DNA, the polymerase extends primers with free dNTPs, forming a complementary second strand.
Applications & Downstream Experiments of SensiFAST™ Probe No-ROX Kit, from Bioline
Intended uses of Bioline’s SensiFAST™ Probe No-ROX Kit, from Bioline include end point PCR, as well as related PCR cloning and amplification techniques. The resulting product is applicable for use in sequencing, and other tested methods.
PCR Troubleshooting Tips
Contamination and inappropriate primer construction create amplification inefficiencies. Therefore, PCR cleanup kits enhance the purity of either the starting or final sample, maximizing accuracy, and effectiveness. Dedicated primer design tools may also be useful in those respects. Optimum temperatures suggested by Invitrogen should be enforced, e.g., elongation (stated in the product hanbook). Fidelity and enzyme concentration (see product specifications) also need to be accounted for when setting up your PCR.