Para la recogida sencilla y no invasiva de saliva y la conservación del ADN en muestras de saliva a temperatura ambiente
Detalles
Supporting Data
Figure 1. Stability of DNA Preserved in Norgen's Saliva DNA Preservative at Room Temperature for up to 75 Months.
Saliva samples were collected from numerous donors and mixed, and then an equal volume of Norgen's Saliva DNA Preservative was added to the saliva. The preserved saliva was then stored at room temperature for up to 75 months. Saliva DNA was subsequently isolated at 1 week, 3 weeks, 1 month, 2 months, 4 months, 6 months, 8 months, 12 months, 20 months, 32 months, 40 months, 52 months and 75 months from 0.5 mL of the saliva/preservative sample using Norgen's Saliva DNA Isolation Kit (Cat. RU45400). For visual analysis, 100 µL of the purified DNA was run on an agarose TAE gel. As it can be seen, there is no evidence of DNA degradation after the saliva samples are stored for 75 months at ambient temperatures in Norgen's Saliva DNA Preservative. Furthermore, the size of the DNA was maintained over 24 kb for the entire 75-month period. M: Norgen's UltraRanger 1 Kb DNA Ladder (Cat. 12100).
Figure 2. GAPDH Real Time PCR Amplification of Saliva DNA from Saliva Samples Stored at Room Temperature from Day 0 up to 24 Months.
All isolated saliva DNA samples were successfully amplified without any sign of PCR inhibition and difference in Ct. Red: Day 0, Blue: 4 months, Purple: 8 months, Green: 16 months and Sky blue: 24 months. Black: No template control.
Figure 3. V3-V4 16s rRNA PCR amplification for Illumina MiSeq 16s rRNA library preparation.
The saliva DNA was isolated using Norgen's Saliva DNA Isolation Kit (Cat. RU45400) from saliva that had been preserved for various periods of time up to 2 years at room temperature in Norgen’s Saliva DNA Preservative. The PCR result shows the clean PCR amplification at ~550 bp indicating the right target of V3-V4 16s rRNA , with no difference between day 0 and 24 months. M: FastRunner DNA Ladder (Cat. 12800).
Datos de respaldo
|
Especificaciones del kit
|
|
|
Volumen de saliva recogida
|
2 mL
|
|
Volumen de la mezcla saliva-conservante
|
4 mL
|
|
Temperatura de conservación
|
Temperatura ambiente
|
| Tiempo de conservación |
Más de 2 años a temperatura ambiente
|
Conservación y manipulación
- Cuando se almacenan a temperatura ambiente, los tubos conservantes de saliva de Norgen no utilizados son estables hasta la fecha de caducidad sin ninguna reducción en su rendimiento. Consulte la fecha de caducidad en el prospecto o en la etiqueta del tubo.
- Una vez recogida, la saliva es estable durante más de 2 años si se mantiene herméticamente cerrada y almacenada a temperatura ambiente.
- El tubo de recogida, el embudo de recogida y el recipiente de cada dispositivo individual de recogida y conservación de ADN de saliva son reciclables.