Product Details
Kit Size:
48 Reactions
Product Name:
DNA Isothermal Rapid Amplification Kit Fluorescent Type
Kit Type:
DNA
Reaction Volume:
50 μL
Storage Temperature:
-20℃
Application:
Nucleic Acid Amplification
Type:
Fluorescent Type
Reaction Time:
20mins
High Light:
,
,
Payment & Shipping Terms
Minimum Order Quantity
48T
Price
3.8$/T
Packaging Details
16T/bag,48T/box
Delivery Time
6days
Payment Terms
T/T,Paypal
Supply Ability
100000T/Month
Product Description
Product parameters:
Reagent component ( WLE8202KIT ,16T/bags,48T/Box ) | |||
Component | Specification | Quantity | Function |
A buffer | 1.6ml | 1 Tube | Buffer system mainly for stabilizing protein/enzyme and performance |
B buffer | 0.15ml | 1 Tube | Mainly activated systems such as magnesium ions |
Positive control template | 0.1ml | 1 Tube | Mainly the positive plasmid template is used to test the effectiveness of the kit |
Positive control primer mix | 0.06ml | 1 Tube | Mainly the primer combination of the positive control template |
Reagent Guide Manua | 16T/bags,48T/Box | 3 bags | Reagent technology of protein/enzyme system: freeze-dried powder, freeze-dried microspheres |
Principle overview
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
Primer design
It is recommended to use primers with a length of 30-35 bp. Primers that are too short will affect amplification speed and detection sensitivity; primers are designed to avoid the formation of secondary structures that affect amplification; the amplicon length is recommended to be 150-300 bp, usually no more than 500 bp.
Fluorescent probe design
The probe sequence does not overlap with the specific primer recognition site, is 46-52 nt in length, and the sequence avoids palindromic sequences, internal secondary structures, and continuous repeated bases. The probe has four modification sites: the middle position ≥ 35 nt from the 5′ end is labeled with a dSpacer (tetrahydrofuran, THF) as the recognition site for exonuclease; the upstream of the THF site is labeled with a fluorescent group, and the downstream Label a quenching group, the distance between the two groups is 2-4 nt; THF is ≥15 nt from the 3′ end, and the 3′ end is labeled with a modifying group, such as an amine group, a phosphate group or a C3-Spacer.
Product features and advantages:
This kit has the advantages of high sensitivity, strong specificity,and short reaction time (only 20 minutes), and the reaction groups are in dry powder state, which is easy to operate and easy to store.
It can be applied to various brands of fluorescence quantitative PCR instruments, constant temperature fluorescence amplification instruments and other fluorescence detection equipment.
This kit is based on a room temperature and constant temperature nucleic acid rapid amplification technology: at room temperature and constant temperature, the recombinase and primer form the protein/single-stranded nucleotide complex Rec/ssDNA, with the help of auxiliary proteins and single-stranded binding protein SSB , invade the double-stranded DNA template; form a D-loop region at the invasion site, and start scanning the DNA double-strands; after finding the target region complementary to the primer, the Rec/ssDNA complex disintegrates, and the polymerase also binds to The 3′ end of the primer initiates chain extension. This kit relies on the action of exonuclease at 39 ºC, adding specific molecular probes designed based on the template, and using fluorescence monitoring equipment to achieve real-time monitoring of the amplification process of the target fragment.
RNA/DNA/Protein Purification Plus Kit
This kit provides a rapid method for the high throughput isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, or yeast. The kit employs two columns: 1) for gDNA purification and 2) for RNA purification utilizing Norgen’s resin columns (superior for the binding of all RNA sizes including miRNA). The proteins are also purified on the second column after RNA elution. The proteins are eluted in buffer and are ready for downstream application without any further clean up required. The proteins can be quantified directly, used in western blots, ELISA or mass spectrometry. This kit provides a rapid spin-column method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants.
RNA/DNA/Protein Purification Plus Micro Kit
This kit provides a rapid spin-column method for the isolation and purification of total RNA, DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, microdissected samples including LCM, stem cells, sorted cells, and CTC. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes. The RNA and DNA can be eluted in as little as 20 µL while the protein can be eluted in as little as 50 µL. This kit provides the same performance as if the samples were isolated from dedicated kits.
RNA/DNA/Protein Purification 96-Well Plus Kit
The kit employs two plates: 1) for DNA purification and 2) for RNA purification utilizing Norgen’s resin (superior for the binding of all RNA sizes including miRNA). Please see the protocol schematic below.
Figure 1 / 4
Click for expanded view
Kit Specifications | |
Binding Capacity Per Well | 50 μg for RNA 20 μg for DNA 150 μg for protein |
Size of RNA Purified | All sizes, including small RNA (< 200 nt) |
Size of DNA Purified | ≥ 30 kb |
Time to Complete 10 Purifications | 35 minutes |
Maximum Amount of Starting Material:Animal CellsAnimal TissuesBloodBacteriaYeastFungiPlant Tissues | 1 x 106 cells10 mg100 µL1 x 109 cells1 x 108 cells40 mg40 mg |
Average Yield* Liver (5 mg) | 12.5 μg RNA 2 μg DNa 55 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).
Component | Cat. 47700 (50 preps) | Cat. 51600 (50 preps) | Cat. 51700 (96 preps) |
---|---|---|---|
Buffer SKP | 40 mL | 40 mL | – |
Lysis Buffer Q | – | – | 40 mL |
Wash Solution A | 2 x 38 mL 1 x 18 mL | 2 x 38 mL 1 x 18 mL | 2 x 38 mL 1 x 18 mL |
Elution Solution A | 6 mL | 6 mL | 20 mL |
Elution Buffer F | 15 mL | 15 mL | 2 x 15 mL |
Wash Solution C | 30 mL | 30 mL | 60 mL |
Binding Buffer A | 8 mL | 8 mL | 8 mL |
Elution Buffer C | 8 mL | 8 mL | 30 mL |
Protein Neutralizer | 4 mL | 4 mL | 4 mL |
Protein Loading Dye | 2 mL | 2 mL | 3 x 2 mL |
gDNA Purification Columns | 50 | – | – |
gDNA Purification Micro Columns | – | 50 | – |
gDNA Purification 96-Well Plate | – | – | 1 |
RNA/Protein Purification Columns | 50 | – | – |
RNA/Protein Purification Micro Columns | – | 50 | – |
RNA/Protein Purification 96-Well Plate | – | – | 1 |
Collection Tubes | 150 | 150 | – |
Collection Plate | – | – | 5 |
Elution Tubes (1.7 mL) | 150 | 150 | – |
Elution Plate | – | – | 3 |
Lysis Preparation Plate | – | – | 2 |
Adhesive Tape | – | – | 4 |
Product Insert | 1 | 1 | 1 |
HiPure Gel Pure DNA 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 >100bp from agarose gel(<0.5g), purification of DNA from PCR, enzymatic reaction solution or crude gDNA |
Applications | PCR, NGS, labeling, ligation and enzyme digestion, etc. |
Purification method | Mini spin column |
Purification technology | Silica technology |
Process method | Manual (centrifugation or vacuum) |
Sample type | Agarose gel, PCR products, enzyme products |
Sample amount | Agarose gel: ≤500mg |
Recovery | ≥80% |
Elution volume | ≥15μl |
Time per run | ≤20 minutes(1-24 samples) |
Liquid carrying volume per column | 800µl |
Binding yield of column | 35µg |
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 | D211102 | D211103 |
Purification Times | 100 Preps | 250 Preps |
Buffer GDP | 120 ml | 250 ml |
Buffer DW2 | 50 ml | 2 x 50 ml |
Elution Buffer | 20 ml | 30 ml |
HiPure DNA Mini Columns II | 100 | 250 |
2 ml Collection Tubes | 100 | 250 |
Storage and Stability
The Kit should 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.
Experiment Data
HiPure Gel Pure DNA 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.
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