FP210-HRM Analysis Kit (EvaGreen)

HRM Analysis Kit(EvaGreen)
——Professional reagent for high resolution melting curve analysis

HRM Analysis Kit (EvaGreen)
Cat. No. 4992776/4992873

Storage
The HRM Analysis Kit (EvaGreen) can be stored at -30~-15°C for one year.
It should be stored immediately after receipt at -30~-15°C, protected from
light. Thaw the 2× HRM Analysis PreMix and 50× ROX Reference Dye and
mix thoroughly before use. If the 2× HRM Analysis PreMix and 50× ROX
Reference Dye are thawed but not used, it is important to thoroughly mix
them prior to refreezing. The delamination of salts during the thawing
process and subsequent crystallization during freezing will damage the
enzyme and decrease product performance. For frequent use, HRM
Analysis PreMix can be stored at 2-8°Cfor 3 months. Repeated freeze-thaw
cycles should be avoided.

Introduction
HRM analysis Kit (EvaGreen) combines the advantages of saturation dye EvaGreen
and antibody modified enzyme, and is specially designed to perform High
Resolution Melting (HRM) analysis. A 2×pre-mixed solution is included in this
kit which greatly simplifies the preparation of qPCR reaction mixture. It is highly
sensitive and could be used to analyze different types of templates.
Unlike SYBR Green, high concentration of EvaGreen dye will not inhibit PCR
reaction, but could increase dsDNA products binding, so it is called “saturation
dye”. In contrast with SYBR Green, “dye rearrangement” will not happen when
using EvaGreen, so it could properly distinguish the difference on single base of
amplification products.
This Kit can be used in known SNP analysis, unknown mutant gene scan and
Methylation Specific PCR (MS-PCR).

Features

1. HRM Analysis PreMix uses antibody modified hot start DNA polymerase, along
   with the specially optimized buffer solution, provides a high sensitive and high
   efficient analysis method with wide sample range.
2. HRM Mix includes EvaGreen saturation dye, which has high melting resolution
   under saturation condition, so it could detect the mutation of single base.
3. The unique buffer system in this kit strengthens the stability of melting curve,
   increasing the specificity of amplification.
4. This Kit includes ROX Reference Dye, which can eliminate the interference of
   signal background and revise the deviation of fluorescence signal between
   different wells.

Principle of Operation
SYBR Green is the most widely used fluorescent dye in Real Time PCR, but its
concentration need to be very low in the PCR reaction mixture since it has
inhibition effect toward the PCR reaction. The binding condition of SYBR Green
and dsDNA is not saturated, so it is called “unsaturation dye”. This trait of SYBR
Green will lead to the “dye rearrangement” effect, and then influence the
resolution of melting curve. In this condition the difference between single bases
cannot be distinguished by analyzing melting curve when using SYBR Green as
fluorescent dye.

EvaGreen is a novel fluorescent dye which can be used both in Real Time
PCR and HRM analysis. This dye can selectively bind to dsDNA through
a specific mechanism called “release according to requirement”. This
mechanism not only maximally reduces the PCR inhibition effect of
fluorescent dye, but also allows the HRM analysis to detect single base
differences when the fluorescent dye is under saturation condition

FP210-HRM Analysis Kit (EvaGreen)

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