GenTegra® Frequently Asked Questions.
What are the two options for stabilizing RNA in GenTegra-RNA?
Option 1 – Following application to GenTegra-RNA, use RNA in liquid form for up to 100 hrs on ice or at room temperature (21-25°C). GenTegra-RNA conveys additional stability to RNA in liquid form by inactivating trace RNase activity, simplifying sample handling. Following the 100 hour period, dry the sample down, or store RNA according to your typical protocol.
Option 2 – Following application to GenTegra-RNA dry RNA solution and store or transport at ambient temperature. Following rehydration, GenTegra-RNA conveys additional stability to RNA in liquid form for up to 8 hours at room temperature (21-25°C) or on ice. Following the 8 hour post-recovery period, dry your sample down again for storage or store according to your typical protocol.
Where is the GenTegra-RNA and how can I detect it?
The GenTegra-RNA Cluster Tubes, 0.5 ml Tubes, or 96- well microplates may appear to be empty. The GenTegra-RNA is supplied as a coating at the bottom of each GenTegra-RNA Tube or well, which may appear as a white coating. To confirm that the kit you received contains the GenTegra-RNA, simply rehydrate one tube with 35 μL of molecular biology grade water and take an absorbance reading at 230nm to detect the GenTegra-RNA.
What is GenTegra-RNA? Is GenTegra-RNA composed of a filter, beads or paper?
GenTegra-RNA is not a filter, beads or paper. GenTegra-RNA is a water soluble, inert chemical matrix.
Where can I purchase new caps for GenTegra-RNA screw cap tubes? Are looped/tethered caps available?
New caps may be purchased from any lab supply company. Axygen microcentrifuge tube screw caps are compatible with GenTegra-RNA tubes and are available through several distributors: Genesee Scientific offers the caps in a variety of colors and styles, including looped/tethered caps, at: www.geneseesci.com.
Can RNA be rehydrated and dried multiple times?
Yes, RNA may be rehydrated and dried up to five times, If the sample needs to be used more often than five times it should be aliquoted into tubes for a single additional rehydration.
Can I use the 5X bulk to make custom tubes?
We recommend using the 1X concentration so a larger surface area at the bottom of the tube is coated with GenTegra-RNA but it is just as acceptable to deposit 5 μl of 5X in the bottom of the tube.
Is it safe to keep RNA at room temperature (21-25°C) during the 16 hour drying process?
Yes, the GenTegra-RNA protects RNA in the liquid state at room temperature during the drying process.
What is the composition of the storage solution after recovery?
After addition of molecular-grade water, your samples will be in the same buffer they were stored in at the time of application, plus the inert stabilizers in GenTegra-RNA.
How should I store my recovered RNA?
Following recovery, RNA may be stored for up to 8 hours at room temperature (21-25°C), and then dry down or store according to your typical protocol.
Can I use the recovered RNA directly for downstream applications?
Additional purification is not required prior to performing downstream applications. Similar RNA quality is maintained before and after recovery.
Can GenTegra-RNA tubes be used to store DNA?
No, the chemical matrix used to store DNA is not the same as the chemical matrix used to store RNA. Use GenTegra-DNA tubes for storage and transport of purified DNA samples.
What is GenTegra™? Is GenTegra™ composed of a filter, beads or paper?
GenTegra-DNA is not a filter, beads or paper. GenTegra-DNA is an inert chemical matrix.
Where is the GenTegra-DNA and how can I detect it?
The GenTegra-DNA Cluster Tubes, 0.5 ml Tubes, or 96- well microplates appear to be empty. The GenTegra-DNA is supplied as a transparent coating at the bottom of each GenTegra-DNA Tube or well. To confirm that the kit you received contains the GenTegra-DNA, simply rehydrate one tube with 35μL of molecular biology grade water and take an absorbance reading at 230nm to detect the GenTegra-DNA.
Can samples stored in low-EDTA TE, water or other buffers be applied to GenTegra-DNA Tubes?
Yes, refer to the table in the User Guide for a list of storage solutions that are compatible with GenTegra-DNA Tubes.
What is the maximum concentration of DNA that can be applied to GenTegra-DNA Tubes?
There is no maximum concentration for application (note that the maximum concentration for recovery is 200 ng/μl). When applying less than 20 μL of DNA, add water to a final volume of ≥20 μl to ensure complete mixing of the DNA with the GenTegra-DNA. Refer to the tables on pages 14-17 of the DNA User Manual for application volume and mass specifications.
Why is there a minimum recovery volume of 35 μl?
A minimum 35 μl volume is required to rehydrate DNA from all surfaces of the tube or well.
Why is there a maximum recovery concentration of 200ng/μL when recovering or concentrating DNA?
Maximum solubility of DNA in water is achieved when the concentration does not exceed 200 ng/μl. At very high concentration (>200ng/uL) high molecular weight DNA becomes very viscous.
What is the composition of the storage solution after recovery?
After addition of molecular biology water, your samples will be in the same buffer they were stored in at the time of application, plus the inert stabilizers in GenTegra-DNA.
Will the GenTegra-DNA affect my DNA quantitation? Do I need to blank the spectrophotometer with the GenTegra-DNA?
The GenTegra-DNA absorbs at 230 nm. Thus, it will not interfere with readings at Å260 or Å280 and blanking with the GenTegra-DNA is not required.
How should I store my recovered DNA?
We recommend storing recovered DNA at 4°C for short-term use and -20°C for long-term storage. Do not store recovered DNA at room temperature unless it is dried in the presence of GenTegra-DNA.
Can I use the recovered DNA directly for downstream applications?
Purification is not required prior to performing downstream applications. Similar DNA quality is maintained before and after recovery. GenTegra-DNA does not remove nucleases or other contaminants present in the original sample. When concentrating DNA, please be aware that contaminants will be concentrated along with the DNA.
How can I convert RPM to g (rcf)?
Use the correct centrifugation speeds in order to maximize yield and purity. Check your centrifuge setting using the following equation: g(rcf) = 1.12*r*(rpm/1000), where r is the radius of the rotor in mm.
I left my protease out at room temperature. Can I still use it?
Yes, up to 24 hours. Store the protease at 4°C. Refer to page 7 in the GenSolve User Guide for additional storage information.
How do I get a material safety data sheet (MSDS)?
MSDS documents are available by contacting Technical Support at firstname.lastname@example.org.
Where can I find further information about DNA quantitation and setting up a PicoGreen assay?
Refer to PicoGreen Assay Protocol provided by supplier.
Is GenSaver a good substitute for FTA cards
Yes, the GenSaver cards was designed to be a better collection card in every way than the old FTA card. The GenTegra-DNA chemistry that is present on the GenSaver cards protects the DAN in the DBS better than FTA cards and also releases the DNA more easily giving on the average 100% more DNA per extraction.
Can GenSaver punches be used for direct PCR?
Yes, the GenSaver chemistry has no interference with direct PCR of a 1-1.2 mm punch commonly used for direct PCR.
Can the GenCollect card be used for DNA collection?
Using the GenCollect card for the collection of DBS for DNA extraction is acceptable if you do not intend to store the DBS samples on GenCollect cards for the long term. The GenCollect card has no added chemistry for protecting the DNA from high temperatures or for storage periods of time greater than a year.
Can I use the GenCollect card like a Guthrie card?
Yes, the GenCollect card is ideal for application where small molecules are the samples of interest and a MassSpec is the analysis method.