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For example, if the cycling program specifies 45 cycles and an annealing time of 10 seconds, a LightCycler^® 480 System PCR run for gene scanning will last about 75 minutes including 10 minutes pre-incubation time and 15 minutes high resolution melting.The same run will last about 80 minutes on the LightCycler^®96 System, including 10 minutes high resolution melting.", "Language": "en", "Country": "XG", "Code": "Assay Time" }, { "Name": "Principle", "Value": "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 (e.g., gel-based) methods.
In a gene scanning experiment, sample DNA is first amplified via real-time PCR in the presence of LightCycler^® 480 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.

LightCycler^® 480 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 (Chou, Q., et al., 1992, Kellogg, D.E., et al., 1994, Birch, D.E., 1996), and LightCycler^® 480 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 pre-incubation step (95°C, 5 – 10 minutes) before cycling begins.
LightCycler^® 480 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 (e.g., SYBR Green I or ethidium bromide). LightCycler^® 480 High Resolution Melting Dye is not inhibitoric 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 (Wittwer, C.T., et al., 2003, Herrmann, M.G., et al., 2006).", "Language": "en", "Country": "XG", "Code": "Principle" }, { "Name": "Product Characteristics", "Value": "Easy-to-use Reaction Mix (2x conc.) for PCR and high resolution melting using the LightCycler^® 480, or LightCycler^® 96 Real-Time PCR Systems.", "Language": "en", "Country": "XG", "Code": "Product Characteristics" }, { "Name": "Content", "Value": "

Vial / Bottle	Cap	Label	Function/Description	Content
1	green	LightCycler® 480 High Resolution Melting Master Mix, 2x conc.	Contains FastStart Taq DNA Polymerase, reaction buffer, dNTP mix (with dUTP instead of dTTP), and High Resolution Melting Dye.	5 vials, 1 ml each
2	blue	LightCycler® 480 High Resolution Melting Master, MgCl2, 25 mM	To adjust MgCl2 concentration.	2 vials, 1 ml each
3	colorless	LightCycler® 480 High Resolution Melting Master, Water, PCR Grade	To adjust the final reaction volume.	5 vials, 1 ml each

", "Language": "en", "Country": "XG", "Code": "Content" }, { "Name": "Applications", "Value": "The LightCycler^® 480 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^® 480 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.", "Language": "en", "Country": "XG", "Code": "Applications" }, { "Name": "Product Description", "Value": "The LightCycler^® 480 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^® 480 High Resolution Melting Master can be used in conjunction with LightCycler^® Uracil-DNA Glycosylase for carry-over prevention during PCR.", "Language": "en", "Country": "XG", "Code": "Product Description" } ] } } ] }