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High-Resolution Melting (HRM)

MEDIBENA Life Science & Diagnostic Solutions

5 x HOT FIREPol® EvaGreen® HRM Mix

With
ROX

 Applied Biosystems 7900HT, 7500, 7700

Without
ROX

 Bio-Rad  CFX384™, CFX96™ (software update Jun 2010) ,Qiagen  Rotor-Gene® Q, Rotor-Gene® 6000  

Tab. 01. Mix compatibility table (Intercalating dye based approach)

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Description

HOT FIREPol® EvaGreen® HRM Mix is an optimized ready-to-use solution for High Resolution Melt (HRM) Analysis, incorporating EvaGreen® dye. It includes: HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2 and ROX dye according to system requirements.

Features

• Sensitive HRM genotyping
• Unique stability at room temperature
• Based on EvaGreen® dye
• Convenient ready-to-use solution
• Reaction set-up without using ice


 

I am very satisfied with the results of the RT-qPCR I made with the 5 × HOT FIREPol® EvaGreen® qPCR Mix Plus (ROX). It's sensitive to low amount of template and works well with Stratagene Mx3005P™ thermocycler I use.
Saad Nizar, PhD Student, University of Strasbourg, Institut de Biologie, Moleculaire et Cellulaire 
_____

Performance Data of HRM Mix

Sensitive HRM genotyping

High Resolution Melt analysis was used to genotype a C insertion in BRCA1 gene, a breast cancer susceptibility gene, with HOT FIREPol® EvaGreen® HRM Mix (two graphs above). Reactions were performed on Corbett Rotor-Gene® 6000. Green lines represent wildtypes without an insertion, red lines represent a C insertion and blue line represents a patient with unknown phenotype.

Applications

• Detection and quantification of DNA and cDNA targets
• Profiling gene expression
• Genotyping
• Microbial detection
• Viral load determination
• High Resolution Melt Analysis

Quality control

• Functional quality control via real-time PCR on different genomic templates
• Tests for stability at room temperature
• Tests for multiple freeze-thaw cycles


   

 

PRODUCT CONTENTS (PDF)
CAT. NO. REACTIONS
5 x HOT FIREPol® EvaGreen®
HRM Mix (ROX
)
HOT FIREPol® DNA Polymerase,
5 x EvaGreen® qPCR buffer, dNTPs,
12.5 mM MgCl2, ROX dye, BSA

08-33-0000S
08-33-00001
08-33-00020
 50
250
5000
5 x HOT FIREPol® EvaGreen®
HRM Mix
 HOT FIREPol® DNA Polymerase,
5 x EvaGreen® qPCR buffer, dNTPs,
12.5 mM MgCl2, BSA
08-31-0000S
08-31-00001
08-31-00020

50
250
5000

Tab. 02. Product table

MEDIBENA Life Science & Diagnostic Solutions

 

 

<h1 style="text-align: left;">Mix compatibility table</h1>
<p>&nbsp;</p>
<table style="border: 1px solid #669000;" border="1" width="600" rules="all" height="267">
<tbody>
<tr>
<td>
<p style="text-align: center;">qPCR platforms</p>
</td>
<td>
<p style="text-align: center;">Probe based<br />approach</p>
</td>
<td>
<p style="text-align: center;">Intercalating dye based<br />approach</p>
</td>
</tr>
<tr>
<td style="text-align: center;"><strong>Applied Biosystems</strong> 7900HT, 7500, 7700, 7000, <br />StepOne&trade; &amp; StepOnePlus&trade; <strong><br />Stratagene &nbsp;</strong>Mx3000P&trade; &amp; Mx3005P&trade;</td>
<td>
<p style="text-align: center;"><a href="http://www.medibena.at/catalogsearch/result/?q=5+x+HOT+FIREPol%C2%AE+Probe+qPCR+Mix+Plus+%28ROX%29">5 x HOT FIREPol&reg; <br />Probe qPCR&nbsp;&nbsp; <br />Mix Plus (ROX)</a></p>
</td>
<td>
<p style="text-align: center;"><a href="http://www.medibena.at/catalogsearch/result/?q=5+x+HOT+FIREPol%C2%AE+EvaGreen%C2%AE+qPCR+Mix+Plus+%28ROX%29">5 x HOT FIREPol&reg; <br />EvaGreen&reg;<br />qPCR Mix Plus (ROX)</a></p>
</td>
</tr>
<tr>
<td style="text-align: center;"><strong>Bio-Rad </strong>CFX96&trade; &amp; CFX384&trade;, iQ&trade;5 &amp; MyiQ&trade;, <br />Chromo4&trade;, Opticon&reg; 2 &amp; MiniOpticon&reg;; <br /><strong>Qiagen </strong>Rotor-Gene&reg; Q, Rotor-Gene&reg; 6000 <br /> <strong>Eppendorf</strong> Mastercycler&reg; <em>ep realplex</em><sup>2</sup> &amp; <em>ep realplex</em><sup>4</sup></td>
<td>
<p style="text-align: center;"><a href="http://www.medibena.at/catalogsearch/result/?q=5+x+HOT+FIREPol%C2%AE+Probe+qPCR++Mix+Plus+%28no+ROX%29">5 x HOT FIREPol&reg; <br />Probe qPCR<br />Mix Plus (no ROX)</a></p>
</td>
</tr>
</tbody>
</table>

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  1. SB_08-31-GP HOT FIREPol EvaGreen® HRM Mix (NO ROX)

    HOT FIREPol EvaGreen® HRM Mix (NO ROX)

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    5 x HOT FIREPol® EvaGreen® HRM Mix is an optimised ready-to-use solution for High Resolution Melt (HRM) Analysis, incorporating EvaGreen® dye. It comprises all the components necessary to perform qPCR and HRM Analysis: HOT FIREPol® DNA Polymerase, ultrapure dNTPs, MgCl2 and EvaGreen® dye. The user simply needs to add water, template and primers. HOT FIREPol® DNA Polymerase is activated by a 15 min incubation step at 95°C. This prevents extension of nonspecifically annealed primers and primer-dimers formed at low temperatures during qPCR setup.

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MEDIBENA Life Science & Diagnostic Solutions | Zeillergasse 38, 1170 Vienna, Austria, Europe | office@medibena.at | TEL: +43-699-1133-0065 | FAX: +43-1-2533-0339-577 | Skype: medibena | Register of companies: Handelsgericht Wien Österreich | FN: 392173a | VAT: ATU67954225 | EORI: ATEOS1000086649 | Dr. Monika Bencurova MEDIBENA Life Science E.U. | www.MEDIBENA.at