B0711 - PureBind Blood Genomic DNA Isolation Kit - Nucleic Acid Isolation Kits - By Applications
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B0711

Details

High Molecular Weight Genomic DNA: High yields of high molecular weight genomic DNA isolated from whole blood with Ocean NanoTech’s Blood gDNA kit. 3 x 350 µL of whole blood/donor was processed through the Ocean NanoTech kit and eluted in 100 µL. 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 Whole Blood, no RNase: High purity genomic DNA isolated from whole blood with Ocean NanoTech’s Blood gDNA kit. 3 x 350 µL of whole blood/donor was 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.

Inhibitor-free genomic DNA from whole blood: SPUD assay analysis of isolated genomic DNA demonstrates absence of co-purified inhibitors. 3 x 350 µL of whole blood/donor was processed through the Ocean NanoTech kit and eluted in 100 µL. 100 ng of eluted DNA were assayed for the ability to inhibit amplification of the SPUD amplicon as detailed in Analytical Biochemistry, 2006 Apr 15;351(2):308-10. Uniformity of the observed Cq with isolated DNA templates relative to the SPUD positive control indicates that the isolated DNA is free of PCR/qPCR inhibitors.

Successful qPCR amplification of genomic targets: qPCR analysis of isolated genomic DNA demonstrates successful amplification and detection of genomic target. 3 x 350 µL of whole blood/donor was processed through the Ocean NanoTech kit and eluted in 100 µL. A nominal 20 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 102%.

Enzymatic modification of isolated genomic DNA: Restriction digestion of genomic DNA isolated from whole blood with Ocean NanoTech’s Blood gDNA kit. 3 x 350 µL of whole blood/donor was processed through the Ocean NanoTech kit and eluted in 100 µL. 1 µg of purified DNA was digested for 30 minutes at 37C with BamHI, BglII and BamHI+BglII. Thereafter the restriction product was electrophoresed on a 1% agarose gel/TAE for 1 hour.

Long range PCR from isolated genomic & mitochondrial DNA: Long range PCR of genomic DNA isolated from whole blood with Ocean NanoTech’s Blood gDNA kit. 3 x 350 µL of whole blood/donor was processed through the Ocean NanoTech kit and eluted in 100 µL. 20 ng of purified DNA was amplified via PCR using TaKaRa LA Taq® and primers for (A) a 7.3 kb fragment of human glucocerebrosidase and (B) 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 blood stabilizer tubes: The Blood Genomic DNA Kit can be used to isolate inhibitor-free gDNA from commercial blood stabilizer tubes.. Donor blood was drawn into 3 commercially available blood stabilization products and stored at room temperature for 4 days. 350 µL aliquots from the tubes were processed through the Ocean NanoTech kit using the standard protocol. The DNA/RNA Shield™ tubes were also processed without the Ocean NanoTech Lysis Buffer as DNA/RNA Shield™ lyses cells as part of the stabilization process. 100 ng of eluted DNA from each isolation were assayed for the ability to inhibit amplification of the SPUD amplicon as detailed in Analytical Biochemistry, 2006 Apr 15;351(2):308-10. Uniformity of the observed Cq with isolated DNA templates relative to the SPUD positive control indicates that the isolated DNA is free of PCR/qPCR inhibitors.

Performance vs. other commercial product: Ocean NanoTech’s Blood gDNA kit significantly outperforms other magnetic bead-based blood gDNA isolation kits. Blood from the same donor was drawn into K2-EDTA tubes and processed through each kit using the kit’s requisite input blood volume. Isolated DNA was eluted in 100 µL and assessed for purity via NanoDrop analysis and by Qubit analysis for concentration. Total yield (based on the Qubit measurements) and the yield as a function of blood input volume were calculated for each kit. All values are derived from three biological replicates.

Catalog# Product Unit Size Unit Price Quantity
B0711-096 PureBind Blood Genomic DNA Isolation Kit 1 µm $199.00
B0711-384 PureBind Blood Genomic DNA Isolation Kit 10 nm $677.00
Product Description PureBind Blood Genomic DNA Isolation Kit
Application Nucleic Acid Isolation from Whole Blood