What sample types can be used with the Inhibitor- Tolerant qPCR Mixes? We have tested whole human and bovine blood, saliva, sputum, urine, cerebrospinal fluid (CSF), cow milk, stool sample, and plant lysate; other types of samples can be used but we would recommend testing different concentrations to determine the concentrations that can be tolerated. Do I need to add additional magnesium to the reaction? The SensiFAST™ Probe Direct SuperMix has been optimized for magnesium, dNTPs, stabilizers, and enhancers, resulting in highly reproducible, accurate RNA and DNA target amplification under fast thermal cycling conditions, delivering excellent results in qPCR multiplex assays, even in the presence of difficult PCR inhibitors. So, we do not recommend adding any additional material, only the primers, probe(s), and sample. SensiFAST TM Probe Direct SuperMix
PRODUCT FAQs
What are the recommended reaction conditions for qPCR multiplex assays? When multiplexing, standard conditions can be used, however, if necessary, the reverse transcription reaction time can be extended up to 20 minutes and/or the temperature can be increased up to 55° C and the annealing/extension time can be extended up to 60 seconds and/or the temperature can be increased up to 65° C, but this will need to be tested during optimization. Where do I begin with optimization of crude samples or crude extracts? We would recommend using a range of crude sample concentrations or extract volumes to ascertain at what point unacceptable PCR inhibition starts to take place. This will vary as some sample types, such as blood and stool specimens typically contain more inhibitors for qPCR/RT- qPCR reactions.
SimpliFi HS Mix Optimised for NGS library amplification with low GC bias
SimpliFi HS Mix is a high-fidelity polymerase mix using aptamer-based hot-start technology, highly suited to amplification of DNA from crude samples, for fast, inexpensive target enrichment, NGS library amplification and cloning applications.
Key Features: • Robust: optimized enzyme/buffer mix promotes reliable amplification of a broad range of targets, including complex DNA extracted from human, animal and plant samples • Specific: the aptamer-based hot-start remains completely inactive during PCR set-up and after amplification, to prevent non-specific products • Higher yield and sensitivity: high yields with minimal optimization, regardless of a template’s GC content, reducing time to results and eliminating the cost of unnecessary repeats • Enhanced accuracy: higher than 130x Taq fidelity, reducing errors for next generation sequencing (NGS) library amplification. High fidelity SimpliFi HS Mix is a combination of the latest advances in buffer chemistry and PCR enhancers and stabilizers, together with an aptamer-mediated hot-start polymerase, dNTPs and MgCl 2 . It has been designed for highly reproducible, accurate assay results in the presence of inhibitors. The advanced buffer chemistry and enhancers has been developed for fast PCR and is designed for superior sensitivity and specificity, making SimpliFi HS Mix perfect for NGS library amplification.
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