用于简单、无创地采集唾液并在室温下保存唾液样本中的 RNA。
细节
Supporting Data
Figure 1. Stability of RNA Preserved in Norgen's Saliva RNA Preservative at room temperature for up to 2 months.
Saliva samples were collected from numerous donors and mixed, and then an equal volume of Norgen's Saliva RNA Preservative was added to the saliva. The preserved saliva was then stored at room temperature for up to 2 months. Saliva RNA was subsequently isolated at day 0, day 4, day 6, day 16, 1 month and 2 months from 0.25 mL of the saliva/preservative sample using supplementary protocol associated with Norgen's Total RNA Purification Kit (Cat. 17200). For visual analysis, 7.5 µL from 50 µL elution was mixed with RNA loading dye and was run on a 1x MOPS agarose gel. RNA integrity was maintained after the saliva samples were stored up to 2 months at ambient temperatures in Norgen’s Saliva RNA Preservative.
Figure 2. Stability of three house keeping genes over a 2 month period.
GAPDH, Human 18S rRNA and beta actin genes were monitored from saliva samples stored at ambient temperature with Norgen's Saliva RNA Preservative. RNA was isolated from 250 µL of preserved samples on day 0, day 4, day 6, day 10, day 16, day 30 and day 60 using Norgen's Total RNA Purification Kit (Cat. 17200). Next, 4 µL of elution was used in 20 µL of the real-time PCR reaction (SYBR Green I) using CFX96 TouchTM Real-Time PCR Detection System (Bio-Rad). All of the genes were preserved and stable over 2 month period.
Figure 3. Norgen’s Saliva RNA Preservative is able to successfully preserve saliva RNA samples when used outside of the ideal range of 1:1 at room temperature for 3 days.
As the amount of saliva added to 2 mL of Saliva RNA Preservative increased from 0.2 mL to 3 mL, RNase protein and β-actin gene were monitored for RNA stability using one-step RT-PCR. The results indicated that the variation of the saliva input volumes did not significantly influence the RNA gene detection.
Figure 4. Detection of Viral, Bacterial and Human RNA from Preserved Saliva Samples.
Saliva (2 mL) collected from a single healthy donor was mixed with 2 mL of Saliva RNA Preservative. Norgen’s COVID-19 positive control from the 2019-nCoV TaqMan RT-PCR Kit (E/RdRP gene) (Cat. TM67200) was spiked into the preserved saliva along with HeLa cells (1.6 m cells /ml ) to generate a contrived sample. From the contrived sample, total RNA was purified using Norgen’s Total RNA Purification Kit (Cat. 17200) and 5 genes (16s rRNA, COVID-19 E gene, RdRP-P1 and P2 gene and RP) were analyzed by PCR or RT-PCR to detect viral, bacterial and human genes.
支持数据
|
试剂盒规格
|
|
|
采集的唾液量
|
2 mL
|
|
唾液-保存剂混合液体积
|
4 mL
|
|
保存温度
|
室温
|
| 保存时间 |
室温下可保存 2 个月
|
保质期和处理
- 在室温下储存时，未使用的 Norgen 唾液保存剂试管可稳定地保存至保质期，其性能不会降低。请参阅设备说明书或试管标签上的保质期。
- 唾液 RNA 采集后，在室温下密封保存可稳定长达 2 个月。
- 每个唾液 RNA 采集和保存装置的采集管、采集漏斗和装置容器均可回收。