High Molecular Weight Genomic DNA: High yields of high molecular weight genomic DNA isolated from saliva with Ocean NanoTech’s Saliva gDNA kit. 6 x 1 mL of collected saliva was processed through the Ocean NanoTech kit and eluted in 100 µL. 3 x 200 µL saliva from the same donor was processed through the optional ‘no wash’ protocol. 500 nanograms of purified DNA was electrophoresed on a 1% agarose gel/TAE alongside the GeneRuler™1kb Plus ladder (Thermo Scientific); uppermost DNA ladder band is 20,000 bp.
High Purity Genomic DNA from Saliva: High purity genomic DNA isolated from with Ocean NanoTech’s Saliva Genomic DNA kit. Cellular pellets from 6 x 1 mL of saliva were processed through the Ocean NanoTech kit and eluted in 100 µL. Eluted DNA was analyzed by spectrophotometric analysis to determine purity ratios and DNA concentration.
Successful qPCR amplification of genomic targets: qPCR analysis of isolated genomic DNA demonstrates successful amplification and detection of genomic target. Cellular pellets from 6 x 1 mL of saliva were processed through the Ocean NanoTech Saliva Genomic DNA kit and eluted in 100 µL. A nominal 10 ng of eluted DNA (based on Nanodrop analysis) were assayed with a ABI TaqMan™ β-Actin qPCR assay. The standard curve spanned 100 – 0.032 ng (27,600 – 8.8 genome equivalents) of human DNA. Assay efficiency calculated to be 82%.
Long range PCR - demonstration of mitochondrial DNA co-isolation: Long range PCR of genomic DNA isolated from saliva with Ocean NanoTech’s Saliva gDNA kit. Cellular pellets from 2 x 1 mL of saliva were processed through the Ocean NanoTech kit and eluted in 100 µL. 20 ng of purified DNA were amplified via PCR using TaKaRa LA Taq® and primers for a 8.2 kb fragment of mitochondrial NADH-ubiquinone oxidoreductase chain 1 (MT-ND1). Thereafter the PCR products were electrophoresed on a 1% agarose gel/TAE for 1 hour. Successful PCR amplification of these high molecular weight targets highlights the structural integrity of the isolated DNA and our kit’s capacity to isolate mitochondrial DNA.
Compatibility with commercial Saliva DNA stabilizer tubes: The Saliva Genomic DNA Kit can be used to isolate gDNA from commercial saliva stabilizer tubes.. Saliva was collected in commercially available saliva DNA stabilization products and stored at room temperature for 3 days. 400 µL aliquots from the tubes were processed through the Ocean NanoTech kit ± proteinase K without addition of digestion buffer as both stabilization tubes lyse cells as part of the stabilization process. Eluted DNA from each isolation was quantified via Qubit™.