The RNAok™ RNase Inhibitor is a recombinant mammalian RNase inhibitor which possesses very high affinity for eukaryotic pancreatic-type ribonuclease. The RNAok™ RNase Inhibitor forms a 1:1 complex with pancreatic RNase A by noncovalent binding, presenting a noncompetitive inhibitory activity on these pancreatic enzymes. RNAok™ RNase Inhibitor is active against RNase A, RNase B, RNase C but not RNAse H, RNase I, RNase T1, RNase T2, and S1 nuclease. RNAok™ RNase Inhibitor is compatible with RT-PCR enzymes such as AMV, M-MLV and ExcelRT™ Reverse Transcriptase or Taq DNA polymerase.
Detail
Description
The RNAok™ RNase Inhibitor is a recombinant mammalian RNase inhibitor which possesses very high affinity for eukaryotic pancreatic-type ribonuclease. The RNAok™ RNase Inhibitor forms a 1:1 complex with pancreatic RNase A by noncovalent binding, presenting a noncompetitive inhibitory activity on these pancreatic enzymes. RNAok™ RNase Inhibitor is active against RNase A, RNase B, RNase C but not RNAse H, RNase I, RNase T1, RNase T2, and S1 nuclease. RNAok™ RNase Inhibitor is compatible with RT-PCR enzymes such as AMV, M-MLV and ExcelRT™ Reverse Transcriptase or Taq DNA polymerase.
Application
cDNA synthesis
in vitro translation
in vitro transcription
One-step RT-PCR
Separation and identification of specific ribonuclease activities
Storage Buffer
40 mM HEPES-KOH (pH 7.5), 100 mM KCl, 8 mM DTT, 0.1 mM EDTA, stabilizer and 50% (v/v) glycerol
Storage
-20°C for 24 months
Other Products
HiDi® 2x PCR Master Mix
Product Info
Document
Product Info
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 therefor, is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).HiDi® 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.This PCR mix is also available with a full-length Taq DNA polymerase with a nuclease domain, featuring 100% compatibility with hydrolysis probes (TaqMan® probes etc.).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® 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.It has also been shown that HiDi® DNA polymerase family is highly suitable for quality control in CRISPR-Cas or TALEN-based applications.Several independently conducted studies show that HiDi® 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.
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.
Document
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 therefor, is the first choice for applications that rely on this property such as allele-specific PCR (asPCR) that is also termed allele-specific amplification (ASA).
HiDi® 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.
Sequentially purify total RNA and total proteins from a single sample
Kit includes a gDNA elimination column
No sample splitting required
No phenol step required for efficient isolation
Ideal for small or difficult to obtain samples
Purify RNA and proteins from cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants
Rapid and efficient spin column procedure
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
This kit provides a rapid, single column method for the isolation and purification of total RNA (including miRNA) and proteins sequentially from a single sample of cultured animal cells, tissues, blood, bacteria, yeast, fungi or plants. The total RNA and proteins are both column purified in under 25 minutes using a single column.
Purified RNA is of a high quality and yield, and is suitable for NGS, RT-qPCR and microarrays.
This kit eliminates DNA efficiently using a gDNA removal column.
Proteins are eluted in buffer and are ready for downstream applications such as Western Blots, Mass Spec and ELISA. The proteins will not require precipitation, resuspending of pellets, or any further cleaning.
This kit is ideal for researchers who are interested in studying the transcriptome and proteome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen’s RNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.
Average Yields*: HEK 293 Cells (1 x 106 cells) HEK 293 Cells (1 x 106 cells) Liver (15 mg) Liver (15 mg)
10-15 μg RNA 70-100 μg protein 30-35 μg RNA 100-150 μg protein
* Average yields will vary depending upon a number of factors including species, growth conditions used and developmental stage.
Storage Conditions and Product Stability The Protein Loading Dye should be stored at -20°C after the addition of DL-Dithiothreitol (DTT). All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.
