N-(Acid-PEG4)-N-bis(PEG4-Propargyl) is a trifunctional PEG linker that combines a t-butyl ester with two terminal alkynes. The alkynes can be applied in copper-click chemistry with azides to form stable triazole linkages with a target molecule, while the acid can be reacted with amines or alcohols to form stable amides or esters.
N-(Acid-PEG4)-N-bis(PEG4-Propargyl) is a trifunctional PEG linker that combines a t-butyl ester with two terminal alkynes. The alkynes can be applied in copper-click chemistry with azides to form stable triazole linkages with a target molecule, while the acid can be reacted with amines or alcohols to form stable amides or esters.
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.
Specifications
| Features | Specifications |
| Main Functions | Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column |
| Applications | RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation |
| Purification method | Mini spin column |
| Purification technology | Silica technology |
| Process method | Manual (centrifugation or vacuum) |
| Sample type | Animal tissues, adherent cells, suspension cells, bacteria, etc |
| Sample amount | Eukaryotic culture cells: ≤ 10^7, Animal tissue:<100mg, Yeast culture cells:<5 x10^7, Bacteria:<10^9 |
| Elution volume | ≥15μl |
| Time per run | ≤40 minutes |
| Liquid carrying volume per column | 800µl |
| Binding yield of column | 100µg |
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such asguanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein andother impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
This kit combines acid/guanidine (MagZol) extraction technology with glass fiber filter membrane purification, which can improve the extraction effect of complex samples and samples with low RNA content. After the sample is treated with MagZol reagent and chloroform, the supernatant is added with ethanol to provide appropriate binding conditions, then transferred to the purification column and centrifuged. Macromolecular RNA can be efficiently bound to the membrane. Collect the filtrate containing small RNA, add more ethanol to adjust the binding capacity of small RNA, the pollutants can be efficiently washed away by second cleaning. Finally, the purified RNA was eluted by RNase free water.
Advantages
Kit Contents
| Contents | R431002 | R431003 |
| Purification Times | 50 Preps | 250 Preps |
| HiPure RNA Mini Columns | 100 | 2 x 250 |
| 2ml Collection Tubes | 100 | 2 x 250 |
| MagZol Reagent | 60 ml | 270 ml |
| Buffer RWC | 20 ml | 80 ml |
| Buffer RW2* | 20 ml | 2 x 50 ml |
| RNase Free Water | 10 ml | 30 ml |
Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.
Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (<57°C)
– Pipetting aid in form of a blue dye for better visibility during aliquoting into well-plates
– included lysis function*
* = A sample extraction and sample lysis is not necessary as immediate hot PCR cycling conditions can brake cell- and virus membranes. Please see our published references to this product.
Note: Volcano3G® RT-PCR Probe 2x Master Mix is a unique master mix. If you are using it for the first time, we recommend to design primers to have higher melting/annealing temperatures (above 65°C). Additionally, for assay setup it is advised to perform RT-PCR test reactions with temperature gradients (i.e., for the annealing and extension steps) to identify the most proficient PCR protocol for your needs using our Volcano DNA polymerase products.
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.
Volcano3G® RT-PCR Probe 2x Master Mix contains all components required for general RT-qPCR
– optimized reaction mix for sensitive and reliable results
– engineered, truly thermostable Taq DNA polymerases with reverse transcriptase activity
– fast start function due to a hotstart aptamer formulation which prevents unspecific amplification at lower temperatures (
Norgen’s EXTRAClean Urine Cell-Free Circulating RNA Purification Kits provide a fast, reliable, reproducible, and simple procedure for isolating circulating RNA and exosomal RNA from various urine inputs ranging from 250 μL and up to 30 mL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extensions, and expression array assays.
EXTRAClean Cell-Free Circulating RNA Purification Mini Kit:
For sample volumes ranging from 250 μL to 2 mL
EXTRAClean Cell-Free Circulating RNA Purification Midi Kit:
For sample volumes ranging from 2mL to 10mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL
EXTRAClean Cell-Free Circulating RNA Purification Maxi Kit:
For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
Figure 1 / 3
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| Kit Specifications | |
|---|---|
| Sample Type | Fresh or frozen urine |
| Sample Volume Range | 250 μL to 2mL |
| Minimum Elution Volume | 50 μL |
| Maximum Elution Volume | 100 μL |
| Time to Complete 10 Purifications | 25–30 minutes |
| Size of RNA Purified | All sizes, including miRNA and small RNA (< 200 nt) |
| Average Yields ¥ | Variable depending on specimen |
¥Please check page 5 of protocol for average urine yields and common RNA quantification methods.
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Component | Mini (Cat. 72900) | Midi (Cat. 72910) | Maxi (Cat. 72920) |
|---|---|---|---|
| Binding Solution K | 25 mL | 75 mL | 1 x 75 mL 1 x 25 mL |
| Lysis Buffer A | 30 mL | 20 mL | 20 mL |
| Wash Solution A | 18 mL | 18 mL | 18 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Collection Tubes | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| EXTRAClean Mini Spin Columns | 50 | 20 | 10 |
| EXTRAClean Midi Spin Columns | – | 20 | – |
| EXTRAClean Maxi Spin Columns | – | – | 10 |
| Product Insert | 1 | 1 | 1 |
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