Saliva collection and preservation gDNA Extraction Kit(SpinColumn)-Genomic DNA Extraction-IBAC

Saliva collection and preservation gDNA Extraction Kit(SpinColumn)

Packing Specification：

D2528A-10	Saliva collection and preservation gDNA Extraction Kit(SpinColumn)	10T	CNY450
D2528A-50	Saliva collection and preservation gDNA Extraction Kit(SpinColumn)	50T	CNY1650
D2528B-50	Saliva gDNA Extraction Kit(SpinColumn)	50T	CNY900
D2528B-100	Saliva gDNA Extraction Kit(SpinColumn)	100T	CNY1600

Product Introduction：
Traditional collection of human genomic DNA samples is usually performed by blood drawing followed by whole blood genomic DNA extraction. This method has several distinct disadvantages: it requires dedicated blood collection equipment and must be conducted by medically trained personnel; the pain of phlebotomy often leads to subjects refusing sampling; invasive collection increases the risk of infection; and blood samples must be transported and stored under low-temperature conditions after collection.
This kit provides a painless, non-invasive method that allows patients to obtain high-quality, high-yield DNA samples without the pain and infection risks of blood drawing. It shows high acceptance among subjects and enables convenient DNA sampling for both infants and the elderly. The collection procedure is very simple: subjects spit saliva into the preservation solution and mix well to complete sample collection. Once mixed, the sample can be transported and stored at room temperature for up to one year without degradation, greatly reducing costs for cold-chain transportation, refrigeration equipment and power consumption. DNA can be extracted from collected saliva via several simple steps, with an average yield of 110 μg from 2 mL saliva.

Product Features：
1.Non-invasive sampling eliminates the pain of blood drawing, lowers contamination risks, and improves sampling convenience, supporting self-collection by subjects.
2.Approximately 110 μg of DNA can be obtained from only 2 mL saliva sample (yields vary widely among individuals).
3.Collected specimens can be stably stored at room temperature for more than one year.

Applications：
1.Collection, preservation, transportation and purification of DNA from saliva samples.

2.Used as an alternative or supplement to Oragene saliva collection tubes.

Storage Conditions：

1.Precipitation may occur in the cell lysis buffer at low temperatures. It can be re-dissolved in a 37 ℃ water bath for several minutes. The buffer can be used after cooling to room temperature once it becomes clear and transparent.
Avoid prolonged exposure of reagents to air to prevent volatilization, oxidation and pH drift. Cap all solutions tightly immediately after use.

Protocol：
Saliva Sample Collection
1.Rinse your mouth with clean water 1–2 times and spit out the water.
2.Wait at least 5 minutes after rinsing before saliva collection. Do not eat or drink any beverages during this period.
3.Spit saliva (not throat mucus/phlegm) into a 5 mL collection tube up to the 2 mL graduation mark.(Do not spit mucus into the collection tube. If saliva volume is insufficient, perform oral movements to stimulate secretion. A small amount of foam floating on the saliva surface is not counted toward the 2 mL target volume. The entire collection must be completed within 30 minutes.)
4.Add the full volume of 2 mL preservation solution into the 5 mL saliva collection tube, mix thoroughly by repeated inversion, then tightly cap the tube.

Saliva DNA Extraction(Using 2 mL saliva as an example; the saliva volume can be scaled up or down proportionally)
Cell Lysis
1.Incubate the preservation solution/saliva mixture in a 50 °C water bath for at least 1 hour, or in a 50 °C air incubator for at least 2 hours.
2.Transfer 4 mL of the mixture (2 mL saliva + 2 mL preservation solution) to a 15 mL or 50 mL centrifuge tube. Add 1 mL Lysis Buffer and 10 μL RNase A solution (10 mg/mL). Vortex vigorously for 10 seconds, then incubate at room temperature for 10 minutes.

Impurity Precipitation
1.Add 1.7 mL Impurity Precipitation Buffer to the lysate mixture above. Vortex vigorously for 25 seconds to fully mix the Impurity Precipitation Buffer with the lysate.
2.Centrifuge at 8,000 × g for 5 minutes. Precipitated impurities and proteins will form a compact pellet at the tube bottom. If the protein pellet is not compact, place the tube on ice for 5 minutes and repeat Step 4.

DNA Precipitation
1.Carefully transfer the supernatant (containing DNA) to a new 15 mL or 50 mL centrifuge tube. Avoid disturbing the pellet at the bottom. Add 5 mL isopropanol.(When DNA content in saliva is low, adding 40 μL Glycogen (20 mg/mL) may slightly improve yield.) Mix gently by inverting 50 times (precipitate may be visible in some cases).
2.Centrifuge at 2,500 × g for 3 minutes. A white DNA pellet will generally be visible at the tube bottom.
3.Discard the supernatant, invert the tube, and tap gently on absorbent paper to drain excess liquid. Add 5 mL 70% ethanol and invert several times to wash the DNA pellet.
4.Centrifuge at 2,500 × g for 1 minute. Carefully discard the supernatant (the pellet is loose; take care not to lose the DNA pellet).
5.Invert the tube and tap gently on absorbent paper to remove residual ethanol. Residual ethanol around the pellet and on the tube wall may also be carefully removed with a pipette tip. Air-dry the pellet for several minutes (do not over-dry, and ensure no ethanol remains).

DNA Hydration & Solubilization
1.Add 250 μL–400 μL DNA Solubilization Buffer to rehydrate and dissolve the DNA pellet; mix by gently flicking the tube wall.
2.Incubate at 65 °C for 30–60 minutes (do not exceed 1 hour), then place at room temperature or 4 °C overnight to fully rehydrate the DNA, with occasional gentle flicking of the tube wall to assist solubilization.
3.DNA may be stored at 2–8 °C. For long-term storage, store at −20 °C or −80 °C.

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