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HotBeads
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Mg HotBead™ For Hot Start PCRThe Mg HotBead™ is a wax bead that releases a precise amount of molecular biology grade MgCl2 into solution when melted. This prevents premature polymerase enzyme activity by withholding magnesium, one of the essential reagents, until the sample is heated above the annealing temperature during the first denaturation step. The non-barrier Mg HotBead™ is recommended for oil-free systems because it will leave a hole in the wax for easy access to the sample. If desired, additional wax can be removed by tipping the tubes over while the wax is melted, spreading the wax onto the sides to leave a larger hole. In an oil-free system, the thermocycler should have a heated cover that minimizes condensation in the tube. If you are using the barrier Mg HotBead™ in place of oil, it will provide a wax barrier over the solution to prevent evaporation. If oil is desired, a drop of oil can be added with the bead, which softens the wax barrier. Procedure Samples are prepared and dispensed as you normally would except that the magnesium is omitted from the master mix. A typical magnesium free, 10X buffer solution consists of: 100 mM Tris-HCl with 500 mM KCl, at pH 8.3. 1. Pour a few beads into a clean weighing boat or flat lid. The Mg HotBead™ is picked up by gently touching it with a plastic pipet tip. Drop it into the tube, close it and start thermocycling. 2. During the denaturation step (1-2 min.), the MgCl2 is automatically released from the bead as the sample is heated above 68 oC. 3. After cooling, the sample is recovered through a hole left in the wax, or with barrier beads, by piercing the wax with a pipet. Suggestions for Using Barrier Beads Before piercing the wax layer, remelt the wax by heating the tubes briefly (one minute @ 80-90 oC), in the thermocycler, then gently tip the tubes over before cooling so that the liquid wax spreads out onto the sides of the tube. This will thin the wax layer over the sample and reduce the chances of plugging the pipet. Also, if desired, piercing the wax is facilitated by using a positive displacement pipet tip. Storage The beads should be stored in the refrigerator. KK Biomed is a licensed distributor for HotBeads. For additional bead sizes and concentrations, please go to HotBead.com . FOR LABORATORY RESEARCH ONLY, NOT FOR DIAGNOSTIC USE. Mg HotBead™ For Hot Start PCRPurchase Orders Welcome by E-Mail or Call: (800) 666-9677
Prices are in US Dollars F.O.B. KK Biomed with the cost of taxes and shipping added to invoice. KK Biomed exclusively warrants that its products perform as described, subject to replacement or refund limited to the purchase price. KK Biomed expressly disclaims warranties of fitness for some other purpose. KK Biomed does not warrant that the use of its products will not infringe the claims of any patents. The purchase of this product does not include a license under any remaining patents that cover the polymerase chain reaction (PCR).
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