Let us build one for your website. Use this tool to find the nucleotide sequence files for commonly used molecular biology tools, including plasmid, viral and bacteriophage vectors. Tm Calculator. The calculator calculates recommended T m (melting temperature) of primers and PCR annealing temperature based on the primer pair sequence, primer concentration, and DNA polymerase used in PCR. The most sophisticated T m calculations take into account the exact sequence and base stacking parameters, not just the base composition(1,2,3).. Our Tm calculator takes actual buffer conditions into account when calculating Tm's for your primer pair by asking you not just for your primer sequences, but for the enzyme and buffer that you intend to use for your reaction. Use this tool to find the right products and protocols for each step (digestion, end modification, ligation and transformation) of your next traditional cloning experiment. NEBCloner Use this tool to find the right products and protocols for each step (digestion, end modification, ligation and transformation) of your next traditional cloning experiment. Simply input your DNA polymerase, primer concentration and your primer sequence and the Tm Calculator will guide you to successful reaction conditions. The Q5 Site-Directed Mutagenesis Kit enables rapid, site-specific mutagenesis of double-stranded plasmid DNA in less than 2 hours (Figure 1). Contact us for more details. NEB offers several helpful interactive tools for your research and experimental design. We use cookies to understand how you use our site and to improve the overall user experience. This functional tool calculates the Tm of primers and estimates an appropriate annealing temperature while using different DNA polymerases. Please check out NEB's Tm Calculator to help you determine your optimal Phusion annealing temperature. NEBcutter V2.0 will indicate cut frequency and methylation state sensitivity. We will post Stimulus Payments as we receive them every business day. Our new RUO kit, the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit, enables simple, visual detection of isothermal amplification of SARS-CoV-2 nucleic acid.. The parameters were adjusted on a set of primers seeking to maximize specificity and retain high yields. Tm value of one of my primer is 52.8 (50mM NaCl) and another one is 54.5 (50mM NaCl) as given by the primer making company. This information is currently unavailable for technical reasons. Use this tool to help select the right DNA polymerase for your PCR setup. Use this tool to interpret Ultra or High Pressure liquid chromatography (UPLC/HPLC) N-glycan profiles following exoglycosidase digestions. This tool designs probes to be used with the NEBNext RNA Depletion Core Reagent Set or to supplement an existing NEBNext Depletion Kit for the depletion of unwanted RNA species. We've developed a NEB online tool, the NEB Tm Calculator, that can be used for any of our polymerases to get an appropriate Ta for your experiment. email or call 1-800-NEB-LABS. Fixed primers can be specified for the design of LAMP primers, and subsequent Loop primers are then designed based on LAMP primer selection. Use the NEB Tm Calculator to estimate an appropriate annealing temperature when using NEB PCR products. Fill out our Technical Support Form, Are you doing COVID-19 related research? Please try again later. If necessary, use a temperature gradient to further optimize and empirically determine the ideal annealing temperature for each of the template-primer pair combinations. NEBuilder Assembly Tool can be used to design primers for NEBuilder HiFi DNA Assembly or Gibson Assembly reactions. 3.8 kb fragment was amplified from 50 ng of Jurkat gDNA using different polymerases. The calculator also calculates the primer length, percentage of GC content, molecular weight, and extinction coefficient. Are you doing COVID-19 related research? Use this tool to guide your reaction buffer selection when setting up double-digests, a common timesaving procedure. NEBuilder Assembly Tool can be used to design primers for your Gibson Assembly reaction, based on the entered fragment sequences and the polymerase being used for amplification. Just fill out your information so we can prioritize what to build. Melting Temperature (Tm) Calculation. Learn about our tools that are helping researchers develop diagnostics and vaccines for the SARS-CoV-2 virus. Manufactured and quality-controlled at New England Biolabs, Thermo Scientific ® Phusion High-Fidelity DNA Polymerase offers both high fidelity and robust performance. A variety of resources to aid you in your work. 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