DBCO-PEG6-DBCO is a monodisperse click chemistry linker containing two terminal DBCO groups with hydrophilic PEG spacer arm. DBCO will react with azide-bearing compounds or biomolecules to form a stable triazole linkage without copper catalyst. PEG spacer arm can increase water solubility and membrane permability. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
DBCO-PEG6-DBCO is a monodisperse click chemistry linker containing two terminal DBCO groups with hydrophilic PEG spacer arm. DBCO will react with azide-bearing compounds or biomolecules to form a stable triazole linkage without copper catalyst. PEG spacer arm can increase water solubility and membrane permability. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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 Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. The extracted products can be used for clinical in vitro detection.
Specifications
| Features | Specifications |
| Main Functions | Isolation circulating DNA from 1-5ml plasma, serum, body fluids using vacuum protocol |
| Applications | qPCR, liquid or solid chip analysis, hybridization and SNP detection, etc. |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (vacuum) |
| Sample type | Serum, plasma and other cell-free fluid samples |
| Sample amount | 1-5ml |
| Elution volume | ≥50μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 4ml |
| Binding yield of column | 1mg |
This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption 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.
| Contents | IVD3182 |
| Purification Times | 50 |
| Buffer ACL | 250 ml |
| Buffer ACB* | 300 ml |
| Buffer DCW1* | 22 ml |
| Buffer DCW2* | 10 ml |
| Proteinase K | 540 mg |
| Protease Dissolve Buffer | 30 ml |
| Carrier RNA | 110 μg |
| Nuclease Free Water | 20 ml |
| HiPure CFDNA Mini Columns | 50 |
| 2 ml Collection Tubes | 100 |
| Extender Tube | 50 |
| Vac-Connector | 50 |
Storage and Stability
Proteinase K, 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 remaining kit 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 should be redissolved before use. Make sure that all buffers are at room temperature when used.
Experiment Data
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,
HiPure DNA Clean Up 96 Kit uses proprietary chemistry and HiPure technology to recover DNA Fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.
Specifications
| Features | Specifications |
| Main Functions | Recover DNA fragments between 60bp-10kbp from PCR, Reactions, crude DNA using 96 well Bind Plate |
| Applications | PCR, sequencing, labeling reactions, ligations and restriction digestion, etc. |
| Purification method | 96 well Bind Plate |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | PCR product, enzymatic reaction |
| Sample amount | Appropriate |
| Recovery | ≥80% |
| Elution volume | ≥75μl |
| Time per run | ≤60 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 20µg |
The HiPure system uses a simple bind-wash-elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.
Advantages
Kit Contents
| Contents | D212201 | D212202 | D212203 |
| Purification Times | 1 x 96 Preps | 4 x 96 Preps | 20 x 96 Preps |
| Buffer GDP | 120 ml | 400 ml | 4 x 400 ml |
| Buffer DW2 | 20 ml | 100 ml | 3 x 100 ml |
| Elution Buffer | 20 ml | 20 ml | 30 ml |
| HiPure DNA Plate | 1 | 4 | 20 |
| 2.2ml Collection Plate | 1 | 4 | 20 |
| 1.6 ml Collection Plate | 1 | 4 | 20 |
| 0.8 ml Collection Plate | 1 | 4 | 20 |
Storage and Stability
The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.
HiPure DNA Clean Up 96 Kit uses proprietary chemistry and HiPure technology to recover DNA Fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.
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.
