DBCO-PEG4-triethoxysilane is a PEG linker containing a triethoxysilane moiety and a DBCO group. Triethoxysilane is commonly used for surface modifications. DBCO group can react with azide-bearing compounds or biomolecules to form a stable triazole linkage without copper catalyst. The hydrophilic PEG chain increasse the water solubility of a compound in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-PEG4-triethoxysilane is a PEG linker containing a triethoxysilane moiety and a DBCO group. Triethoxysilane is commonly used for surface modifications. DBCO group can react with azide-bearing compounds or biomolecules to form a stable triazole linkage without copper catalyst. The hydrophilic PEG chain increasse the water solubility of a compound in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Endonucleases DNA-specific, dsDNase
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
In figure 1, a PCR master mix was treated with different amounts of dsDNase before performing a qPCR to measure the contaminating bacterial DNA in the master mix. ArcticZymes dsDNase effectively removed contaminating DNA below known levels of the assay detection limits.
The dsDNase from Arctic shrimp (Pandalus borealis) is recombinantly produced in Pichia pastoris. It cleaves phosphodiester linkages in DNA to yield oligonucleotides with 5’-phosphate and 3’-hydroxyl termini.
The specific activity is estimated to be 30 times higher than that of bovine DNase I. In the presence of magnesium as only divalent cation and using oligos as a substrate, the activity towards dsDNA is 5000-fold higher than towards ssDNA.
The unique double strand-specificity allows specific degradation of dsDNA while leaving shorter ssDNA as primers and probes essentially intact. Easy inactivation by moderate heat (65°C) allows addition of DNA intended for analysis directly after removal of contaminating DNA.
Figure 1. The dsDNase effectively removes contaminated DNA
The dsDNase effectively removes contaminated DNA:
A PCR master mix was preincubated with various concentrations of dsDNase. After treatment, no DNA was amplified in non-template controls.
Nucleic acid specificity has been tested towards double- and single-stranded DNA and RNA oligonucleotides. The specificity of dsDNase towards the substrate has been measured using 15-mer oligonucleotides with FAM at 5′ and DarkQuencher® 3′ (Eurogentec). The fluorescence is proportional to enzyme activity. Assay conditions: 25 mM Tris pH 7.5, 5 mM MgCl2, and 2 μM oligonucleotide.
Substrate Relative Activity
dsDNA 100%
ssDNA <0.03%
dsRNA <0.01%
ssRNA <0.01%
Double-Strand Specific dsDNase (dsDNase) is ideal for fast and effective removal of contaminating DNA from PCR master mixes.
Taq polymerases are commonly contaminated by bacterial DNA. This is a problem in PCR based bacterial typing and detection as it might cause false positive results. The unique properties of dsDNase make it suited for removal of contaminating DNA from PCR master mixes prior to addition of DNA template.
Introduction
Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments, <1000bp (DNA). HiPure Circulating DNA Mini Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. Samples can be fresh or frozen (provided that they have not been frozen and thawed more than once).
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 1ml plasma, serum, body fluids |
| Applications | qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Serum, plasma and other cell-free fluid samples |
| Sample amount | 1ml |
| Elution volume | ≥30μl |
| Time per run | ≤50 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10 Mm Tris,pH 8.0).
Kit Contents
| Contents | D318102 | D318103 |
| Purification Times | 50 Preps | 250 Preps |
| Buffer ACL | 50 ml | 250 ml |
| Buffer ACB* | 60 ml | 300 ml |
| Buffer DCW1* | 22 ml | 88 ml |
| Buffer DCW2* | 10 ml | 50 ml |
| Proteinase K | 120 mg | 540 mg |
| Protease Dissolve Buffer | 10 ml | 30 ml |
| Carrier RNA | 110 μg | 310 μg |
| Nuclease Free Water | 10 ml | 30 ml |
| HiPure CFDNA Mini Columns | 50 | 250 |
| 2 ml Collection Tubes | 100 | 500 |
Storage and Stability
Proteinase K and carrier RNA should be stored at 2–8°C upon arrival. However, short-term storage(up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remainingkit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions.The entire kit can be stored at 2–8°C, but in this case buffers shouldbe redissolved before use. Make sure that all buffers are at room temperature when used.
Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasma usually as short fragments,
Parvovirus B19 was the first known human virus in the family of Parvoviruses. B19 is a non-enveloped, icosahedral virus that contains a single-stranded linear DNA genome. It is classified as an erythrovirus because of its capability to invade red blood cell precursors in the bone marrow. B19 virus causes a childhood rash called fifth disease or erythema infectiosum which is commonly called “slapped cheek” syndrome due to the appearance of erythema across the cheeks. The virus is primarily spread by infected respiratory droplets, but some cases of blood-borne transmission have been reported. Parvovirus infection in pregnant women is associated with hydrops fetalis due to severe fetal anemia, sometimes leading to miscarriage or stillbirth. The risk of fetal loss is about 10% if infection occurs before week 20 of pregnancy. There is also some evidence that intrauterine Parvovirus B19 infection leads to developmental abnormalities in childhood. Due to the serious nature of the virus, methods for its rapid diagnosis are desirable.
Parvo B19 TaqMan PCR Kit, 100 reactions
Parvo B19 TaqMan PCR Probe/Primer Set and Controls, 100 reactions
Figure 1 / 3
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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.
| Component | Cat. TM39650 (100 preps) | Cat. TM39610 (100 preps) |
|---|---|---|
| MDx TaqMan 2X PCR Master Mix | 2 x 700 μL | – |
| Parvovirus B19 Primer & Probe Mix | 280 μL | 280 μL |
| Parvovirus B19 Positive Control | 150 μL | 150 μL |
| Nuclease-Free Water (Negative Control) | 1.25 mL | 1.25 mL |
| Product Insert | 1 | 1 |
