For life science research only. Not for use in diagnostic procedures. Others LightCycler High Resolution Melting Master LSR LightCycler® High Resolution Melting Master 3.5.8.1.1.4 04 909 631 001 4 909 631 001 04909631001 4909631001 04909631001 LightCycler HRM Master LightCycler High Resolution Melting Master 04038377024837 Reagents, kits 5 x 1 mL 2x conc.5 x 100 reactions of 20 μL final volume each false The LightCycler® High Resolution Melting Master is designed for life science research studies in combination with the LightCycler® 480, or LightCycler® 96 Real-Time PCR System. The LightCycler® High Resolution Melting Master is a ready-to-use 2x conc. hot start reaction mix designed for amplification and detection of a specific DNA sequence (if suitable PCR primers are provided) followed by high resolution melting curve analysis for detection of sequence variants among several samples. A separate 25 mM MgCl2 stock solution, supplied with the Master, allows you to easily optimize the Mg2+ concentration. en The LightCycler® High Resolution Melting Master is designed for life science research studies in combination with the LightCycler® 480, or LightCycler® 96 Real-Time PCR System. It is supplied as a ready-to-use 2x concentrated hot start reaction mix. A separate 25 mM MgCl2 Stock Solution, supplied with the master, allows users to easily optimize the Mg2+ concentration. The LightCycler® High Resolution Melting Master can be used in conjunction with LightCycler® Uracil-DNA Glycosylase for carryover prevention during PCR. en Gene scanning or mutation scanning techniques detect the presence of sequence variations in target-gene-derived PCR amplicons. The method is based on high resolution melting, a novel, closed-tube post-PCR method enabling genomic researchers to analyze genetic variations in PCR amplicons prior to or as an alternative to sequencing. High resolution melting provides high specificity, sensitivity, and convenience at significantly higher speed and much lower cost than other established methods, such as gel-based methods. In a gene scanning experiment, sample DNA is first amplified via real-time PCR in the presence of LightCycler® High Resolution Melting Dye. After PCR, the successive melting experiment can be performed on the same instrument, and analyzed with the LightCycler® 480, or LightCycler® 96 Software to identify sequence variants. Thus, the entire experiment can be done on the instrument without opening the reaction vessels and without additional handling steps after the PCR setup. How this product works LightCycler® High Resolution Melting Master is a ready-to-use reaction mix developed for the detection of DNA samples that differ in sequence from others. It contains FastStart Taq DNA Polymerase for hot start PCR, which significantly improves the specificity and sensitivity of PCR by minimizing the formation of nonspecific amplification products, and LightCycler® High Resolution Melting Dye for monitoring of formation and denaturation of double-stranded DNA. FastStart Taq DNA Polymerase is a chemically modified form of thermostable recombinant Taq DNA Polymerase* that shows no activity at temperatures up to +75°C. The enzyme is active only at high temperatures, where primers do not bind nonspecifically. The enzyme is activated (by removal of blocking groups) during a single preincubation step (+95°C, 5 to 10 minutes) before cycling begins. LightCycler® High Resolution Melting Dye is a new fluorescent dye that enables detection of sequence variations by different melting curve shape, especially if a sample is heterozygous for a particular mutation. This feature is not shared with other dyes traditionally used in real-time PCR, such as SYBR Green I or ethidium bromide. LightCycler® High Resolution Melting Dye is not inhibiting to amplification enzymes. Thus, high concentrations of the dye do not affect the PCR. These high concentrations completely saturate the dsDNA in the sample. dsDNA remains dye saturated during the subsequent melting experiment. Under these conditions, even single nucleotide exchanges result in subtle, but reproducibly detectable changes in melting curve shape. en

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LightCycler^® High Resolution Melting Master

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