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.
Barrier Beads Vs. Non-Barrier Beads
 | Both types of beads provide a wax vapor barrier during thermal cycling. |
| However, 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. |
| The Non-Barrier Beads are engineered to provide a small hole in the wax after cooling, for immediate sample access. |
Application
| Mg2+ HotBeads® can be used with all polymerases that require MgCl2. |
| Use HotBeads® for a uniform vapor barrier in place of messy paraffin oil |
| HotBeads® give equivalent HotStart results to other methods |
Instructions
| Prepare master mix without MgCl2 |
| Aliquot required volumes to each tube |
| Add a bead to each tube |
| Start cycling. |
Important Tips
| Beads are easily dispensed from a weighing boat by touching with a disposable pipette tip. |
| After 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. |
| Seal the tubes again for later analysis or storage by re-melting and cooling vertically. |
Other Information
|
melting point 68-74 C |
|
shape: spherical |
|
nucleases none detected |
|
composition: paraffin, m.b.g. MgCl2 and water. |
© KK Biomed 2007