Frequently asked questions from our customers
Product related Questions: Snooplex
The Taq polymerase is extremely temperature stable thus we are able to ship the Snooplex FastPrep kit at room temperature. We recommend storing in the freezer after receipt.
It depends on the size of your target band. For medium and large targets between 300 and 1000 bp, we recommend control mix A (231 bp). For small targets (100-250 bp), control mix B should be used. If no internal control is desire, please use control mix C.
If you want to purify the PCR product and use it further, e.g. for DNA sequencing, we recommend to completely omit the control mix and to replace the missing volume in the PCR reaction by PCR water instead. Any gel loading buffer can be mixed with an aliquot of PCR product before gel electrophoresis.
The Snooplex lysis buffer is routinely used at GVG GM for DNA extraction from all types of mouse and rat sample material. It was found that the conditions originally recommended for swabs (30 minutes at 70°C) do not work for tail biopsies, for example. In a series of experiments it was investigated whether it is possible to define lysis conditions that are equally suitable for all samples. This is the case. We use incubation at 90°C for 60 minutes, with gentle shaking (500 rpm).
Important: Not too much sample material should be used for DNA extraction. 1-3 mm from a tail tip or 1 - 2 ear punches are completely sufficient to perform up to 50 PCR runs with 2 µl DNA extract each under optimized PCR conditions. The essential prerequisite for this is the use of the Snooplex PCR chemistry and the PCR conditions optimized for low DNA quantities (primer quantity).
If no control band or your PCR products are visible at all, too little DNA has been added. Either sampling was insufficient (the swab needs to be rotated three times) or no DNA has been added to the PCR mix.
Important: You have to ensure that sufficient cell material is collected via swabbing. Sample collection is a vital step and must therefore be carried out properly. Note that you are collecting cell material from mucosa and not just a saliva sample.
In order to handle a competitive situation regarding the amplification of the internal mouse control and your primers, the Snooplex control primers are set up in such a way that PCR products will be produced in favor of your primers, and 231 bp (or 850 bp) control band might fade out.
Yes, the reaction buffer is ready-to-use.
Important: For any PCR, there is always an optimal ratio between the number of DNA molecules (targets) and the amount of primers. The amount of DNA from swabs is considerably lower compared to tail biopsies. Therefore, the originally validated standard PCR conditions are logically suboptimal for low DNA amounts and should be adjusted. The two most important parameters to improve PCR conditions are the number of PCR cycles and the amount of PCR primers used. Therefore, use a DNA dilution series to test your standard conditions and adjust them if necessary. As a good orientation, how far you can dilute the DNA, serves the internal PCR control in the control mix A or B. As long as this band is still visible, there is a sufficient amount of DNA for a PCR. For this, increase the number of PCR cycles to 40. Usually, the amount of target-specific PCR primers added to the reaction can be reduced by 50% or even more. By the way, the optimal PCR conditions found for small amounts of DNA also work wonderfully for larger amounts of DNA.
We strongly recommend to use Snooplex PCR chemistry because it is optimized for small DNA amounts. In-house PCR reagents might fail under these conditions.
Yes, this is possible. Each of the components can also be purchased individually, or defined components can be excluded from the delivery.
For a quick and easy calculation of the composition of the mastermixes our "Snooplex Mastermix-Calculator" can be used. This is provided free of charge.