A common mistake amongst xTAG® users occurs when the wrong buffer is used at the wrong step. You may ask yourself “why did Luminex make it so complicated?” as you rend your garments and gnash your teeth in frustration. But please remain calm, as the answer to your question is simple: Each buffer is specifically designed to work in conjunction with a particular enzyme utilized during a specific reaction. Let me explain this a little more.
For example, the xTAG® RVP kit utilizes the xTAG® 5X OneStep Buffer and the TaKaRa 10X RT PCR Buffer*. Each buffer contains specific amounts of salts and cofactors that allow the corresponding enzymes to function properly. The xTAG® 5X OneStep Buffer is designed to be used with the xTAG® OneStep Enzyme Mix. It is provided at a 5X concentration so that when it is used with the Multiplex RT-PCR master mix recipe, it will be diluted to an ideal concentration. Likewise, when the TaKaRa 10X RT-PCR Buffer is used with the TaKaRa Taq® HS Polymerase for the Multiplex TSPE master mix, the two work in conjunction to promote enzyme activity and are essential for detecting the respiratory virus targets.
Now that you have relaxed and we have clarified the reason for multiple buffers, let me upset you again and remind you that there is still one more buffer included in the xTAG® RVP Kit. This final buffer has traditionally been the xTAG® 10X Buffer and is not involved with any enzymatic activity. I will now give you a moment to stop waving your fist at me. So here’s why: the xTAG® 10X Buffer is diluted to a 1X concentration and is then used to prepare the Streptavidin-Phycoerythrin (SA-PE) reporter. While it does not affect any enzymes, the xTAG® buffer is designed to ensure that Luminex’s beads remain suspended properly and optimally interact with the Luminex 200’s optics system.
Thankfully, we have heard your complaints and have updated this component of the kit to make your lives a bit simpler. Starting June 11th, 2012 Luminex has replaced the xTAG® 10X Buffer in the xTAG® RVP kit with xTAG® Reporter Buffer (First lots affected are: Lot # IK019C-0048, Abbott Lot # IK019-0045, please refer to the embedded Customer Advisory Notices for your region** for additional information). xTAG® Reporter Buffer is a pre-diluted, ready-to-use version of the xTAG® 10X Buffer, removing the need to manually dilute before using. If you have any questions about this change or would like to thank us for this change personally, please contact your Field Applications Specialist or Luminex’s Technical Support Team at 1-877-785-BEAD(2323) or [email protected].
* Takara Taq® HS Polymerase and buffer are not provided in the xTAG RVP® kit and must be purchased from the manufacturer or an approved vendor.
** [download id=”2″] is for US customers; [download id=”1″] is for Canadian Customers. This change does not affect customers outside of US and Canada.