Storage | Purification | Downstream Applications

Storage

Q.	What temperature should I store my saliva samples in Oragene•RNA?
A.	Oragene•RNA samples can be stored at room temperature for up to 8 weeks or at -20°C indefinitely. The RNA will remain stable under these storage conditions. We do not recommend storing the samples at 4°C.

Purification

Q.	What is the RNA yield?
A.	In a study of 16 randomly-selected samples, the average total RNA yield per 2 mL of saliva is 23.4 µg. The range was 10.0 to 53.0 µg.

Q.	What is the best way to quantify my RNA?
A.	RNA that has been purified according to our standard purification protocol can be quantified by absorbance or by using a fluorescent method such as RiboGreen.

Q.	The Agilent Bioanalyzer reports that the RNA extracted from my Oragene RNA/saliva samples is degraded/poor quality. Why is this?
A.	We do not recommend using the bioanalyzer to assess quality of RNA from saliva. Saliva is not a sterile source and the RNA will be both human and bacterial. The mixture of RNA types can not be differentiated by the bioanalyzer and will result in misleading assessment of quality.

Downstream Applications

Q.	Do you have a list of genes that are known to be expressed in cells found in saliva?
A.	The following genes have been found to be expressed in saliva: ß-2-microglobulin, Interleukin-8, Histatin-3, 18S Ribosomal RNA, ß-actinBAG1, CDC37L1, FKBP4, FKBP5, HSP70, HSP90, NCOA1, NCOR1, NR3C1, PPIA, PPID, PPP5C, P23, STIP1, STUB1, ST13, TFRC. Please note that is not an exhaustive list. You will need to empirically determine if the message for your gene-of-interest is found in saliva.

Q.	What is the best way to quantify my RNA?
A.	RNA that has been purified according to our standard purification protocol can be quantified by absorbance or by using a fluorescent method such as RiboGreen.

Q.	The Agilent Bioanalyzer reports that the RNA extracted from my Oragene RNA/saliva samples is degraded/poor quality. Why is this?
A.	We do not recommend using the bioanalyzer to assess quality of RNA from saliva. Saliva is not a sterile source and the RNA will be both human and bacterial. The mixture of RNA types can not be differentiated by the bioanalyzer and will result in misleading assessment of quality.

Q.	How much of my extracted RNA should I use in my reverse transcription reaction?
A.	We recommend using 6 µL of your purified RNA to generate cDNA. RNA will be both human and bacterial, thus using RNA quantification is not an accurate method for determining your input.

"I found the Oragene•DNA collection kit very simple to use. The protocol was easy to follow and time-saving permitting me to perform the extraction amplification and analysis steps in a single day." Melissa Boweman, Technologist Plexagen Diagnostics Ltd.