| Description | Product Introduction:This kit is based on spin column adsorption technology and is suitable for recovering 50 bp–30 Kb DNA fragments from agarose gels of various concentrations. In addition, the kit is also suitable for recovering and purifying DNA from PCR products, enzymatic reaction Product Introduction:This kit is based on spin column adsorption technology and is suitable for recovering 50 bp–30 Kb DNA fragments from agarose gels of various concentrations. In addition, the kit is also suitable for recovering and purifying DNA from PCR products, enzymatic reaction solutions, or crude DNA (including genomic DNA) obtained by various methods. Buffer PB contains a pH indicator, and the solution is yellow, which facilitates judging whether the pH value of the solution is suitable for binding to the DNA adsorption column. The DNA recovery efficiency can be as high as 80%, and the purified DNA can be directly used for sequencing, ligation, restriction enzyme digestion (enzyme digestion), PCR, labeling, and other applications.Product Components and Storage Conditions: U1492721 ComponentComponentStorage U1492721ABuffer BL30 mlRT U1492721BBuffer PB25 mlRT U1492721CBuffer DW212 mlRT U1492721DBuffer EB10 mlRT U1492721EFineBind MinElute DNA Spin Columns50个RT U1492721F2 ml Collection Tubes50个RTStorage Conditions:This kit can be stored for 12 months at room temperature (15°C–25°C) under dry conditions. Precipitation may form in Buffer PB at low temperatures; before use, it is necessary to redissolve the buffer in a 37°C water bath and shake it well before use.Precautions:1. The addition of Buffer BL can improve the adsorption capacity of the adsorption column, enhance its uniformity and stability, and eliminate the impact of high temperature/humidity or other adverse environmental factors on the adsorption column. Before use, please check whether Buffer BL is turbid. If turbidity occurs, heat it in a 37°C water bath for a few minutes to restore clarity.2. Buffer PB contains a pH indicator and is yellow, indicating a pH ≤ 7.5.Operating Steps:Before use, add absolute ethanol to Buffer DW2. Please refer to the label on the bottle for the volume to be added.I. Recovering DNA Fragments from Agarose Gels1. Column Equilibration Step: Add 500 µl Buffer BL to the adsorption column (FineBind MinElute DNA Spin Columns) placed in a collection tube. Centrifuge at 12,000 rpm for 1 minute, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube. (Please use columns processed on the same day)2. Cut the single target DNA band from the agarose gel (remove excess parts as much as possible) and place it in a clean centrifuge tube, then weigh it.3. Add an equal volume of Buffer PB to the gel block (if the gel weighs 0.1 g, its volume can be regarded as 100 µl, so add 100 µl Buffer PB). Incubate in a 50°C water bath for about 10 minutes, gently inverting the centrifuge tube up and down continuously during this period to ensure the gel block is fully dissolved. (If the volume of the gel block is too large, it can be cut into small pieces in advance.)Note: For recovering small fragments < 150 bp, the volume of Buffer PB can be increased to 3 times to improve the recovery rate; after the gel block is completely dissolved, it is best to cool the solution to room temperature before loading onto the column, because the adsorption column has a stronger ability to bind DNA at room temperature. The gel should appear yellow after complete dissolution, and then subsequent operations can be performed. If the color of the solution is orange-red or purple after the gel is completely dissolved, use 10 µl of 3M sodium acetate (pH 5.0) to adjust the color of the solution to yellow before proceeding with subsequent operations. (Buffer PB contains a pH indicator. When pH ≤ 7.5, the solution is yellow, and DNA can effectively bind to the membrane. When the pH is high, the solution turns orange-red or purple and needs to be adjusted.)4. Add the solution obtained in the previous step to the adsorption column (placed in the collection tube), centrifuge at 12,000 rpm for 30 seconds, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube.Note: The capacity of the adsorption column is 700 µl. If the sample volume is larger than 700 µl, it can be added in batches.5. Add 500 µl Buffer DW2 (with absolute ethanol added before use) to the adsorption column, centrifuge at 12,000 rpm for 30 seconds, discard the waste liquid in the collection tube, and put the adsorption column back into the collection tube.6. Repeat step 5.7. Centrifuge at 12,000 rpm for 3 minutes.8. Place the adsorption column into a clean centrifuge tube, 悬空滴加 an appropriate amount of Buffer EB (Buffer EB is heated at 65°C for 3-5 minutes before use, preheated in advance) to the middle part of the adsorption membrane, and let it stand at room temperature for 1 minute. Centrifuge at 12,000 rpm for 1 minute to collect the DNA solution.Note: The volume of the eluent should not be less than 30 µl; a smaller volume will affect the recovery efficiency. If the downstream experiment is sensitive to pH, sterile water can be used for elution. The pH of the eluent has a great impact on the elution efficiency. If water is used as the eluent, ensure its pH is within 7.0-8.5 (NaOH can be used to adjust the pH of water to this range). The elution efficiency is low when the pH is below 7.0.II. Recovering DNA from PCR Reaction Solutions or Restriction Enzyme Digestion Solutions1. Column Equilibration Step: Add 500 µl of Buffer BL to the adsorption column (FineBind MinElute DNA Spin Columns) placed in a collection tube. Centrifuge at 12,000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube. (Please use columns processed on the same day.)2. Calculate the volume of the PCR reaction solution or restriction enzyme digestion solution, add an equal volume of Buffer PB to it, and mix thoroughly (there is no need to remove paraffin oil or mineral oil).Note: For recovering small fragments < 150 bp, the volume of Buffer PB can be increased to 3 times to improve the recovery rate; after mixing, the solution should appear yellow before proceeding with subsequent operations. If the solution is orange-red or purple, use 10 µl of 3M sodium acetate (pH 5.0) to adjust the color of the solution to yellow before continuing.3. Add the solution obtained in the previous step to the adsorption column (placed in the collection tube), let it stand at room temperature for 2 minutes, centrifuge at 12,000 rpm for 30 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.Note: The capacity of the adsorption column is 700 µl. If the sample volume exceeds 700 µl, add it in batches.4. Add 500 µl of Buffer DW2 (ensure absolute ethanol is added before use) to the adsorption column, centrifuge at 12,000 rpm for 30 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.5. Repeat step 4 once.6.Centrifuge at 12,000 rpm for 3 minutes.7. Transfer the adsorption column to a clean centrifuge tube, suspend and add an appropriate amount of Buffer EB (preheat Buffer EB by heating at 65°C for 3–5 minutes before use) to the middle of the adsorption membrane, and let it stand at room temperature for 1 minute. Centrifuge at 12,000 rpm for 1 minute to collect the DNA solution.Note: The volume of the eluent should not be less than 30 µl; a smaller volume will reduce recovery efficiency. If the downstream experiment is sensitive to pH, sterile water can be used as the eluent. The pH of the eluent has a significant impact on elution efficiency. If water is used as the eluent, ensure its pH is within the range of 7.0–8.5 (NaOH can be used to adjust the pH of water to this range); elution efficiency will be low if the pH is below 7.0... Read More | DescriptionThe 1 µm Coupling Kit makes conducting immunoprecipitation and biomolecule separation easier and more flexible. The Kit contains AnteoBind™activated 1 µm magnetic particles that give you increased antibody binding capacity and functionality, while the included blocking DescriptionThe 1 µm Coupling Kit makes conducting immunoprecipitation and biomolecule separation easier and more flexible. The Kit contains AnteoBind™activated 1 µm magnetic particles that give you increased antibody binding capacity and functionality, while the included blocking buffer decreases background noise. Reduce reagent preparation time; remove traditional surface preparation steps such as EDC and replace these steps with the 1 µm pre-activated magnetic particles provided. This Kit reduces aggregation and gives you the freedom and ability to develop multifunctional particles for diverse applications, including dual labelling.Binding Capacity and Dispersity:Binding Capacity:> 20 µg IgG/mgMonodispersity:> 90% (by light microscopy determination)Particle based immunoassays, bioseparations and immunoprecipitation... Read More | Inquire | This plant protein extraction kit can extract soluble plant proteins from fresh, frozen, or dried plant tissues. Suitable for protein extraction from various plants and different parts of plants (such as roots, stems, leaves, flowers, seeds, etc.), with high extraction efficiency, high protein yieldThis plant protein extraction kit can extract soluble plant proteins from fresh, frozen, or dried plant tissues. Suitable for protein extraction from various plants and different parts of plants (such as roots, stems, leaves, flowers, seeds, etc.), with high extraction efficiency, high protein yield, high activity, and fast speed. The extracted protein can be directly subjected to protein electrophoresis analysis, immunoprecipitation, Western Blot, protein activity determination, and protein purification experiments. The concentration of the extracted protein can be determined using the BCA protein quantification kit. P665757Component100 TStorageP665757APlant Protein Extraction Reagent100 mLRTP665757BProtease Inhibitor Cocktail (100×)1 mL-20℃. Avoid freeze/thaw cycle. Precautions：1. This product contains 1mM EDTA.2. To prevent protein degradation, all operations should be carried out on ice as much as possible.3. After extracting protein using this product, the BCA method can be used for protein quantification.4. To achieve the best experimental results, please adjust the optimal usage amount according to the experiment.Operation steps:1. Please remove the required Plant Protein Extraction Agent for pre cooling before protein extraction.2. Weigh the weight of the experimental plant tissue. Add 5 ml of Plant Protein Extraction Agent to 1 g of tissue (add Protein Inhibitor Cocktail in a 1:99 ratio before protein extraction).Attention:1) Before homogenization, cut large pieces of plant tissue into small pieces and homogenize them with a mechanical homogenizer for 10 seconds, with an interval of 10 seconds. Repeat the process three times and select the appropriate homogenization method according to the different tissue samples.2) The amount of lysate used is adjusted according to different parts of the plant. If concentrated protein extracts are needed, the amount of Plant Protein Extraction Agent used can be appropriately reduced.3. After homogenization, incubate on ice for 20-30 minutes.4.4 ℃ 13400 × g, centrifuge for 20 minutes.5. Collect soluble proteins from the supernatant for further purification or downstream analysis... Read More | This reagent kit is based on TRIzon's improved columnar total RNA extraction kit. This product can be extracted from animal groupsExtract total RNA from samples such as textiles, plant materials, various microorganisms, and cultured cells. Firstly, the cracking solution is fully cracked This reagent kit is based on TRIzon's improved columnar total RNA extraction kit. This product can be extracted from animal groupsExtract total RNA from samples such as textiles, plant materials, various microorganisms, and cultured cells. Firstly, the cracking solution is fully cracked andHomogenized samples, in their unique high salt state, RNA specifically binds to silicon matrix membranes, greatly reducingEffectively removing organic solvent contamination while removing protein contamination, resulting in higher purity and quality of RNA. bookThe product can quickly extract total RNA from various cells or tissues, and can process 30-50 mg of tissue or 5 × 10 ⁶ cells each time,Can handle multiple different samples simultaneously. If it is an RNA experiment that is very sensitive to trace amounts of DNA, the residual DNA can be utilizedUsing DNase without RNase for digestion and removal on the column, the extracted RNA can be directly applied to RT-PCR Experiments such as Northern Blot, Dot Blot, and in vitro translation. U665516 Component 50 T Storage U665516A DNase I 1000 U -20℃. Avoid freeze/thaw cycle. U665516B 10×Reaction Buffer 1000 µL -20℃. Avoid freeze/thaw cycle. U665516C TRIzon Reagent 60 mL 2-8℃. Protect from light. U665516D TRIzon PaI™ 10 mL 2-8℃. Protect from light. U665516E Buffer RW1 40 mL RT U665516F Buffer RW2 (concentrate) 11 mL RT U665516G RNase-Free Water 10 mL RT U665516H Spin Columns RM with Collection Tubes 50 sets RT U665516I RNase-Free Centrifuge Tubes (1.5 mL) 50 EA RTPreparation and important precautions before the experiment:1.To prevent RNase pollution, attention should be paid to the following aspects:1) RNase's plastic products and gun heads to avoid cross contamination.2) Prepare the solution using water without RNase.3) Operators should wear disposable masks and gloves, and change gloves frequently during the experiment.2. The sample should avoid repeated freezing and thawing, otherwise it will affect the yield and quality of RNA extraction.3. If TRIzon Reagent is found to have precipitates before use, it can be dissolved in a water bath at 56 ℃ for a few minutes.Before the first use, anhydrous ethanol should be added to Buffer RW2 according to the instructions on the reagent bottle label.5. All centrifugation steps should be carried out at room temperature unless otherwise specified, and all operation steps should be carried out quickly.Usage：1. Sample processing1a. Organization: 30-50 mg of tissue is thoroughly ground in liquid nitrogen and 1 mL of TRIzon Reagent is added, or 1 mL of TRIzon Reagent is added to the tissue sample and homogenized. Attention: The sample volume should not exceed 10% of the volume of TRIzon Reagent.2a. Single layer cell culture: Remove the culture medium and add an appropriate amount every 10 cm ² Add 1 mL of TRIzon Reagent.3a. Cell suspension: Collect cells by centrifugation. Add 1 mL of TRIzon Reagent to every 5 × 10 µ m cell.2. After adding TRIzon Reagent, repeatedly blow a few times to fully crack the sample. Leave at room temperature for 5 minutes to completely separate the protein nucleic acid complex.3. Add 200 to every 1 mL of TRIzon Reagent µ LTRIzon PaI ™， Cover the tube tightly, vigorously shake for 15 seconds, and let it sit at room temperature for 2 minutes.4. Centrifuge at 4 ℃ 12000 rpm (~13400 × g) for 10 minutes. At this time, the sample is divided into three layers: the red organic phase, the middle layer, and the upper colorless aqueous phase. RNA is mainly in the upper aqueous phase. Move the upper aqueous phase to a new RNase Free centrifuge tube (provided).5. Add an equal volume of 70% ethanol (prepared without RNase water) to the obtained aqueous solution, invert and mix well.6. Add all the solutions obtained in the previous step to the spin columns RM that have been loaded into the collection tube. If the solution cannot be added at once, it can be transferred in multiple batches. Centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.7. Add 350 to the adsorption column µ L Buffer RW1, centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.8. Preparation of DNase I mixture: Take 52 µ LRNase Free Water, add 8 to it µ L 10 x Reaction Buffer and 20 µ L DNase I (1 U/ µ L) Mix well and prepare to a final volume of 80 µ The reaction solution of L.9. Directly add 80 µ L DNase I mixture to the adsorption column and incubate at 20-30 ℃ for 15 minutes.10. Add 350 to the adsorption column µ L Buffer RW1, centrifuge at 12000 rpm for 1 minute, discard the waste liquid, and place the adsorption column back into the recovery manifold.11. Add 500 to the adsorption column µ L Buffer RW2 (check if anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 20 seconds, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.12. Repeat step 11.Centrifuge at 12000 rpm for 2 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes and thoroughly air dry. Attention: The purpose of this step is to remove residual ethanol from the adsorption column, which will affect subsequent enzymatic reactions (enzyme digestion,. )PCR, etc.14. Place the adsorption column in a new RNase free centrifuge tube and add 30-50 to the middle of the adsorption column µ Place RNase Free Water at room temperature for 1 minute, centrifuge at 12000 rpm for 1 minute, collect RNA solution, and store RNA at -70 ℃ to prevent degradation.Attention:1) The volume of RNase Free Water should not be less than 30 µ L. Small volume affects the recovery rate.2) If you want to increase RNA production, you can use 30-50 µ Repeat step 14 for the new RNase Free Water.3) If you want to increase the RNA concentration, you can add the obtained solution back to the adsorption column and repeat step 14... Read More |