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Mg2+ HotBead  Introduction

Hotstart PCR using Mg2+ HotBeads can provide dramatic improvement to your PCR reactions by preventing non-specific products.  

Mg2+ HotBeads work by  with holding the  Mg2+ required for Taq activity.  Mg2+ is released when the template is denatured during the first denature cycle. For difficult templates, hotstart PCR can often make the difference between observing product or no product.

Mg2+ HotBeads are a convenient, inexpensive and simple alternative to other Hot Start methods. 

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Barrier Beads Vs. Non-Barrier Beads

bulletBoth types of beads provide a wax vapor barrier during thermal cycling.
bulletHowever, Barrier Beads provide a wax barrier after cooling in an upright position. The sample can be accessed later by re-melting 1 minute and cooling the tubes tilted on their sides.
bulletThe Non-Barrier Beads are engineered to provide a small hole in the wax after cooling, for immediate sample access.

Application

bulletMg2+ HotBeads can be used with all polymerases that require MgCl2.
bulletUse HotBeads for a uniform vapor barrier in place of messy paraffin oil
bulletHotBeads give equivalent HotStart results to other methods 

Instructions

bulletPrepare master mix without MgCl2
bulletAliquot required volumes to each tube
bulletAdd a bead to each tube
bulletStart cycling.

Important Tips 

bulletBeads are easily dispensed from a weighing boat by touching with a disposable pipette tip. 
bulletAfter thermal cycling, Barrier Beads have a thin  wax seal.  For sample removal, heat the tubes 75-80 C for 1 minute to melt the wax and then tilt the tubes 45-60 degrees to cool. This creates a large hole in the wax seal.
bulletSeal the tubes again for later analysis or storage by re-melting and cooling vertically.

Other Information

bullet

melting point 68-74 C

bullet

shape: spherical

bullet

nucleases none detected

bullet

composition: paraffin, m.b.g. MgCl2 and water.

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         KK Biomed 2007

 

KK Biomed Corporation
          825 N. 300 West, #320
Salt Lake City, Utah 84103
          USA
For more information on KK Biomed, Go To:  http://www.kkbiomed.com 
          
HotBead Information or Comments, E mail: sales@hotbead.com