The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.
Detail
Description
The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.
Features
2’-O-Methyltransferase included for Cap-1 RNA
High capping efficiency
High stability
RNase inhibitor is included to enhance the stability of capping reaction
Application
Generation of 5’Cap-0 (m7Gppp) and Cap-1 (m7GpppNm-) RNA by enzymatic reaction
mRNA synthesis for in vitro translation
Gene expression studies
mRNA vaccine development and therapeutics
Storage
-20°C for 24 months
Other Products
[DS1000] FluoroStain™ DNA Fluorescent Staining Dye (Green, 10,000X), 500 μl
Product Info
Document
Product Info
Description
The FluoroStain™ DNA Fluorescent Staining Dye is designed to be a safer replacement for conventional Ethidium bromide (EtBr) which poses a significant health and safety hazard for its users. The FluoroStain™ DNA Fluorescent Staining Dye offers at least 10 times sensitivity in DNA detection levels, and is capable of detecting double stranded DNA (dsDNA) fragments up to 0.04 ng in electrophoresis analysis. The FluoroStain™ DNA Fluorescent Staining Dye shows a high specificity to the dsDNA, with negligible background signal, making the destaining process entirely optional. FluoroStain™ DNA Fluorescent Staining Dye is compatible with both the conventional ultra violet gel-illuminating systems as well as the less harmful long wave length blue light illumination systems. The emission when bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.
Features:
Excellent for post staining
Sensitivity: 0.04 ng DNA
A safer alternative to EtBr
Compatibility: suitable to blue or UV light
Increased cloning efficiency (blue light)
Storage
Protected from light 4°C for 12 months -20°C for 24 months
Document
The FluoroStain™ DNA Fluorescent Staining Dye is designed to be a safer replacement for conventional Ethidium bromide (EtBr) which poses a significant health and safety hazard for its users. The FluoroStain™ DNA Fluorescent Staining Dye offers at least 10 times sensitivity in DNA detection levels, and is capable of detecting double stranded DNA (dsDNA) fragments up to 0.04 ng in electrophoresis analysis. The FluoroStain™ DNA Fluorescent Staining Dye shows a high specificity to the dsDNA, with negligible background signal, making the destaining process entirely optional. FluoroStain™ DNA Fluorescent Staining Dye is compatible with both the conventional ultra violet gel-illuminating systems as well as the less harmful long wave length blue light illumination systems. The emission when bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.
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.
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads(tRNA Purification) to overcome the hurdle.
The Magnetic Beads(tRNA Purification) solves the tRNA and short DNA/RNA purification problem. The beads with our proprietary technology purify tRNA effectively by removing impurities and unwanted components such as dNTPs, detergents, salts, proteins, and other contaminants. The magnetic bead reagents can be used for oligo (>70 nt) applications.
Workflow without large RNA/DNA contamination
In the case of samples contaminated with RNA/DNA such as rRNA and DNA, our magnetic beads can effectively remove RNA/DNA that are 180 nt and larger. Purified tRNA are ideal for applications requiring high quality, as the fragments are free of impurities and contaminants.
Workflow with large RNA/DNA contamination
Comparison of tRNA and 70 nt oligos recovery. BioDynami beads (tRNA purification) successfully recovered DNA fragments both yeast tRNA and 70 nt oligos.
Recovery of tRNA and 70 nt oligos with BioDynami magnetic Beads (tRNA Purification). Yeast tRNA and 70 nt oligos were used as input. Input and recovered oligos were quantified with ssDNA Quantification kit (BioDynami Cat. # 40043) and RNA Quantification kit (BioDynami Cat. # 40044).
Depletion of larger RNAs. Left panel: depletion of 28S rRNA and 18S rRNA. Right panel: depletion of RNA of 180 nt and larger.
Features
Removal of unwanted components and other impurities
Purification of tRNA and oligos (>70 nt)
tRNA
RNA fragments 70 nt or longer
DNA/RNA hybrid fragments 70 nt or longer
Oligo and chimeric oligo 70 nt or longer
dsDNA fragments 70 bp or longer
ssDNA fragments 70 nt or longer
Removal of larger RNA/DNA contamination:
18S rRNA
28S rRNA
RNA/DNA> 180 nt
Document
Solid Phase Reversible Immobilization magnetic beads has been used for nucleic acids purification because they are effective, simple, and fast. The carboxyl groups coated beads are paramagnetic particles that reversibly bind to nucleic acid. However, there is a drawback for magnetic beads that it can only purify DNA/RNA that are 100 base pairs or longer. tRNA and other DNA/RNA fragments shorter than 100 base pairs are not effectively recovered. We have developed Magnetic Beads (tRNA Purification) to overcome the hurdle.
