Pyruvic Acid Assay Kit

Overview

• Sample collection is non-invasive and painless
• Fast and easy processing using a magnetic bead system
• Isolate high quality genomic DNA
• This kit is also compatible with Norgen’s Saliva DNA Collection and Preservation Devices
• Also available in a 96-well format that can be integrated with a robotic automation system

Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System) provides a fast and reproducible method for isolating genomic DNA from saliva samples collected and preserved using Norgen’s Saliva DNA Collection and Preservation Devices, as well as fresh saliva.  Saliva DNA purified using Norgen’s kit is of the highest quality, and is compatible with a number of downstream research applications including PCR, Southern Blot analysis, sequencing and microarray analysis.

Norgen’s Saliva DNA Isolation Kit (Magnetic Bead System) is also available in a 96-well format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.

Details

Supporting Data

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Kit Specifications
Maximum Saliva Input	0.5 mL preserved saliva 0.25 mL fresh saliva
Average Yield from 0.25 mL of Saliva*	2-5 μg
Average Purity (OD260/280)	1.6 – 1.8
Time to Complete 10 Purifications	30 minutes (Cat. RU55400) 45 minutes (Cat. RU62900)

* Average DNA yield will vary depending on the donor

Storage Conditions and Product Stability
The kit contains ready-to-use Proteinase K which is dissolved in a specially prepared storage buffer. The Proteinase K should be stored at room temperature or 4°C. All other solutions should be kept tightly sealed and stored at room temperature (15 – 25°C). These reagents should remain stable for at least 2 years in their unopened containers.

Component	Cat. RU55400 (50 preps)	Cat. RU62900 (192 preps)
Lysis Buffer F	30 mL	2 x 30 mL
Proteinase K in Storage Buffer	1.2 mL	4 mL
Magnetic Bead Suspension	2 x 1.1 mL	8.5 mL
Elution Buffer B	8 mL	30 mL
Elution Tubes (1.7 mL)	50	–
96-Well Plate	–	2
96-Well Elution Plate	–	2
Adhesive Tape	–	2
Product Insert	1	1

• iDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).HiDi® Taq 2x PCR Master Mix  – ready to use mix simplifies your PCR setup. Only target-specific primers and template need to be added as the mix contains all components for a successful and reliable PCR. This ensures reproducible results, significantly reduces set-up times and the risk of pipetting errors.Please note: This PCR mix is also available as a mix with a nuclease deficient variant, featuring higher robustness towards potential PCR inhibitors and compatibility with real-time dyes such as our GreenDye.Benchmarking with products of competitors conducted by us and others show that the HiDi® DNA polymerase family is the first choice for highly selective PCRs, such as genotyping by allele-specific PCR, HLA genotyping, analysis of single CpG methylation sites or the detection of mutations in a high background of wild-type sequences. By using HiDi® Taq DNA polymerase, less than 10 copies of a mutation can be detected in a background of >10.000 wild-type copies straight away without any other tedious assay optimization.HiDi® Taq DNA polymerase harbours a nuclease function and therefor is also suitable for use with hydrolysis probes (TaqMan® probes etc.). It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control and mutation identification in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® Taq DNA polymerase is ideally suited for use in asPCR in numerous research areas ranging from mutation detection to genome editing. (read more)For research use and further manufacturing.In case you are aiming to use our RUO products as components or for your development of e.g. an IVD medical device, please contact us.

HiDi is available as:

HiDi® DNA Polymerase (>>)
HiDi® Taq DNA Polymerase (>>)
HiDi® 2x PCR Master Mix (>>)
HiDi® Taq 2x PCR Master Mix (>>)

Casestudies:
HiDi® DNA Polymerase: Applications from mutation detection to genome editing (read more)

Example Primer Design

Matching vs. mismatching nucleotide is placed at the 3′-end of the primer for best discrimination results.

Example Results – There´s no accounting for taste

Cilantro: some people love it in their food, some hate it. Here we are detecting a genomic SNP (rs72921001) in HeLa genomic DNA. This SNP is reported to be close to a number of genes coding for olfactory receptors. (Reference: Eriksson N. et al. (2012), “A genetic variant near olfactory receptor genes influences cilantro preference.”)

Considering, that only the C-allele specific primer is extended and yielding in a specific amplicon, we can conclude a genetic predisposition in disliking cilantro, as this SNP is significantly associated with detecting a soapy taste to cilantro.  

Allele-specific PCRs were performed from 1 ng/µl of HeLa gDNA in the presence of a realtime dye, indicating the amplification of the C-allele specific primer only. The A-allele specific primer is discriminated, thus not amplified up to 50 cycles.

PCR products were subsequently analysed on a 2.5% agarose gel. Specific product is visualized by ethidium bromide staining at the amplicon length of 109 bp.

HiDi® stands for High Discrimination of mismatches at the 3’-terminus of primers in PCR. This myPOLS Biotec enzyme family is optimized for this feature and is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) or allele-specific amplification (ASA).