| 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 | Product content: M665559Component50 TStorageM665559ABuffer GTT15 mLRTM665559BBuffer GL15 mLRTM665559CBuffer GW1（concentrate）13 mLRTM665559DBuffer GW2（concentrate）15 mLRTM665559EBuffer GE15 mLRTM665559FProteinase K1.25 mLRTM665559GSpin CoLumns DM with CoLLection Tubes50 Product content: M665559Component50 TStorageM665559ABuffer GTT15 mLRTM665559BBuffer GL15 mLRTM665559CBuffer GW1（concentrate）13 mLRTM665559DBuffer GW2（concentrate）15 mLRTM665559EBuffer GE15 mLRTM665559FProteinase K1.25 mLRTM665559GSpin CoLumns DM with CoLLection Tubes50 EART Product Introduction:This reagent kit is suitable for extracting high-purity total DNA from fresh or frozen mouse or rat tails. The method provided by this reagent kit is simple and feasible, and the purification process does not require phenol or chloroform extraction. It can obtain DNA fragments up to 50 kb, and can also effectively recover fragments of 100 bp. This reagent kit uses a unique lysis solution to effectively lyse mouse tail samples. The optimized buffer system efficiently binds the DNA generated after the lysis of mouse tail to the silica matrix adsorption column, while other pollutants can flow through the membrane; Inhibitors of PCR and other enzymatic reactions can be effectively removed through a two-step washing process, followed by washing with low salt buffer or water to obtain high-purity DNA. The purified DNA can be directly used for downstream experiments such as enzyme digestion, PCR, ReaL Time PCR, library construction, Southern BLot, and molecular labeling.Self prepared reagent: anhydrous ethanol.Preparation and important precautions before the experiment:1. Samples should avoid repeated freeze-thaw cycles, otherwise it may result in smaller extracted DNA fragments and a decrease in extraction volume.2.Before the first use, anhydrous ethanol should be added to BufferGW1 and BufferGW2 according to the instructions on the reagent bottle label.3. Before use, please check if there is any crystallization or precipitation in the Buffer GL. If there is any crystallization or precipitation, please dissolve the Buffer GL again in a 56 ℃ water bath.Operation steps:1. Take a tail of a rat or two mice with a length of 0.4-0.6 cm, grind it into fine powder in liquid nitrogen or cut it into pieces and place it in a centrifuge tube (provided by oneself). Join 180 µ L Buffer GTT, shake and mix well. Note: Ensure that the starting quantity of the organization does not exceed the recommended range.2. Add 20 µ L Protein K, vortex oscillation, thoroughly mix.3. Place in a 56 ℃ water bath until the tissue solution is completely clear. Generally, digestion is required for 6-8 hours. During the incubation process, vortex oscillation is required to evenly disperse the sample. Note: 1) If there is still gel like substance after incubation and vortex oscillation, digest overnight or add 20 more if necessary µ L Protein K digestion will not affect subsequent operations. 2) To remove RNA, add 4 after completing the above steps µ L 100 mg/mL RNase A solution, shake well and let stand at room temperature for 5-10 minutes.4.12000 rpm (~13400 × g) for 1 minute to remove undigested tissues similar to mouse hair. Transfer the supernatant to a new centrifuge tube (provided by oneself).5. Add 200 µ L Buffer GL, vortex oscillation, thoroughly mixed. Join 200 µ L anhydrous ethanol, vortex and shake, thoroughly mix. Short centrifugation allows the solution on the tube wall to be collected to the bottom of the tube.Attention: 1) After adding Buffer GL and anhydrous ethanol, immediately vortex and shake to mix well.2) If multiple samples are operated together, Buffer GL and anhydrous ethanol can be mixed in equal proportions and added to the samples together.3) The addition of Buffer GL and anhydrous ethanol may produce white precipitates, which will not affect subsequent experiments.6. Add all the solutions obtained in step 5 to the adsorption column (Spin CoLumins DM) that has been loaded into the collection tube. If the solution cannot be added at once, it can be transferred multiple times. Centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.7. Add 500 to the adsorption column µ L Buffer GW1 (check if anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.8. Add 500 to the adsorption column µ L Buffer GW2 (check if anhydrous ethanol has been added before use), centrifuge at 12000 rpm for 1 minute, discard the waste liquid in the collection tube, and place the adsorption column back into the collection tube.Note: To further improve DNA purity, repeat step 8.9.12000 rpm for 2 minutes and discard the waste liquid from the collection tube. Place the adsorption column at room temperature for a few minutes to thoroughly air dry. Note: The purpose of this step is to remove residual ethanol from the adsorption column, which will affect subsequent enzymatic reactions (such as enzyme digestion, PCR, etc.).10. Place the adsorption column in a new centrifuge tube (provided by oneself) and add 50-200 to the middle of the adsorption column in the air µ L Buffer GE or sterilized water, leave at room temperature for 2-5 minutes, centrifuge at 12000 rpm for 1 minute, collect DNA solution, and store DNA at -20 ℃.Note: 1) If downstream experiments are sensitive to pH or EDTA, they can be washed off with sterilized water. The pH value of the eluent has a significant impact on the elution efficiency. If water is used as the eluent, its pH value should be ensured to be between 7.0-8.5 (NaOH can be used to adjust the pH value of the water to this range). When the pH value is below 7.0, the elution efficiency is not high.2) Incubating at room temperature for 5 minutes before centrifugation can increase yield.3) Use an additional 50-200 µ Re washing with L Buffer GE or sterilized water can increase yield.4) If you want to increase the final concentration of DNA, you can add the DNA eluent obtained in step 10 back onto the adsorption membrane and repeat step 10; If the elution volume is less than 200 µ L. It is possible to increase the final concentration of DNA, but it may reduce the total yield. If the amount of DNA is less than 1 µ g. Recommended 50 µ L Buffer GE or off... Read More | Products contentN665968Component96 TStorageN665968Adex N501-N508 Primers for Illumina 8×12 µL-20℃. Avoid freeze/thaw cycle.N665968BIndex N701-N712 Primers for Illumina 12×8 µL-20℃. Avoid freeze/thaw cycle. Products IntroductionThis kit is a companion kit for the Products contentN665968Component96 TStorageN665968Adex N501-N508 Primers for Illumina 8×12 µL-20℃. Avoid freeze/thaw cycle.N665968BIndex N701-N712 Primers for Illumina 12×8 µL-20℃. Avoid freeze/thaw cycle. Products IntroductionThis kit is a companion kit for the transposase-based second-generation sequencing Rapid DNA Library Construction Kit, designed for Illumina platform library construction, which contains 8 primers at the N5 end and 12 primers at the N7 end, which can be used to prepare 96 different bipartite Index libraries. All reagents provided in the kit have been subjected to stringent quality control and functional validation to maximize the stability and reproducibility of library construction. The prepared libraries can be sequenced on Illumina platforms such as HiSeq X-10/4000/2500/2000 and MiSeq. Provide your own instruments, reagents and consumables1. Magnetic frame: DynaMagTM-2 is recommended.2. DNA purification and recovery kit: It is recommended to use Kangwei DNA purification and recovery kit by magnetic bead method.3. DNA building kit: It is recommended to use the Kangwei Century transposase method second-generation sequencing rapid DNA building kit.4. Anhydrous ethanol.5. Reaction tubes: It is recommended to use low adsorption PCR tubes and 1.5 ml centrifuge tubes; tips: It is recommended to use high-quality filtration tips to prevent contamination of reagent kits and library samples. Pre-experiment Preparation and Important NotesPlease centrifuge briefly before opening the cap so that the liquid collects at the bottom of the tube to avoid cross-contamination between different primers.For the use of the CombiVision Second Generation Sequencing Multisample Primer Kit, please follow the CombiVision Second Generation Sequencing Rapid DNA Library Kit protocol.Index N501-N508 Primers for IlluminaIndex N701-N712 Primers for Illumina... Read More | Product contentP666142Component200 TStorageP666142ABuffer P160 mLRTP666142BBuffer P260 mLRTP666142CBuffer N380 mLRTP666142DBuffer PB35 mLRTP666142EBuffer PW (concentrate)25 mLRTP666142FBuffer EB30 mLRTP666142GRNase A (10 mg/mL)600 µLRTP666142HSpin Columns DM with Collection Tubes200 EART Product contentP666142Component200 TStorageP666142ABuffer P160 mLRTP666142BBuffer P260 mLRTP666142CBuffer N380 mLRTP666142DBuffer PB35 mLRTP666142EBuffer PW (concentrate)25 mLRTP666142FBuffer EB30 mLRTP666142GRNase A (10 mg/mL)600 µLRTP666142HSpin Columns DM with Collection Tubes200 EART Product IntroductionThis kit is suitable for extracting 1-5 ml of bacterial solution. Based on the lysis of cells by alkaline lysis method, it adopts a unique silica matrix membrane adsorption technology and reagent formulation, and efficiently and exclusively binds plasmid DNA in solution by centrifugal adsorption columns in a high-salt state, and each adsorption column can adsorb a maximum of 30 µg of plasmid DNA, and removes proteins, genomes, RNAs, and other impurities to the greatest extent possible. The plasmid DNA obtained can be directly used for cell transfection, PCR, digestion, sequencing, ligation and other biological experiments.Self-contained reagent: anhydrous ethanol.Pre-experiment Preparation and Important Notes1. All components can be stably stored in dry, room temperature (15-30℃) environment for 1 year, the adsorption column can be stored at 2-8℃ for a longer period of time, and Buffer P1 with RNase A can be stably stored at 2-8℃ for 6 months.2. Before the first use, add all the RNase A solution into Buffer P1, mix well, and store it at 2-8°C. Before use, leave it at room temperature for a period of time, and then use it after recovering to room temperature.3. Anhydrous ethanol should be added to Buffer PW according to the instructions on the label of the reagent bottle before first use.4. If precipitation is found in Buffer P2, Buffer N3, or Buffer PB before use, the clarification can be restored by water bath at 37℃ for a few minutes (please do not shake Buffer P2 violently).5. Be careful not to touch Buffer P2, Buffer N3 and Buffer PB directly, and tighten the lid immediately after use.6. The amount and purity of extracted plasmid is related to the concentration of bacterial culture, strain type, plasmid size, plasmid copy number and other factors.Procedure1. Take 1-5 ml of the overnight culture and add it to a centrifuge tube (self-prepared), centrifuge for 30 seconds at 13,000 rpm (~16,200×g) to collect the bacterial precipitate, and discard the supernatant as much as possible.2. Add 250 µl of Buffer P1 to the centrifuge tube with the bacterial precipitate (please check that RNase A has been added first), mix well using a pipette or vortex shaker, and suspend the bacterial precipitate.Note: If the bacterial mass is not thoroughly mixed, it will affect the lysis effect, resulting in low extraction and purity.3. Add 250µl of Buffer P2 to the centrifuge tube and mix gently up and down 4-6 times, mixing well to lyse the organisms, at which point the solution should become clear and viscous.Note: Mix gently, do not shake vigorously to avoid interrupting the genomic DNA and causing the extracted plasmid to be mixed with genomic DNA fragments. This step should take no more than 5 minutes to avoid damage to the plasmid.4. Add 350 µl of Buffer N3 to the centrifuge tube and immediately mix gently up and down for 8-10 times, mixing well so that a white flocculent precipitate should appear. centrifuge at 13,000 rpm for 5 minutes.Note: Buffer N3 should be mixed immediately after addition to avoid localized precipitation.5. Transfer the supernatant obtained in step 4 to the Spin Columns DM that have been loaded into the collection tube, centrifuge at 13,000 rpm for 30 seconds, pour off the waste liquid from the collection tube, and place the column back into the collection tube.6. Add 150 µl Buffer PB to the adsorption column and centrifuge at 13,000 rpm for 30 seconds.7. Add 400 µl Buffer PW to the adsorption column (please check that anhydrous ethanol has been added first), centrifuge at 13,000 rpm for 1 minute, and pour off the waste liquid in the collection tube.8. Place the adsorbent column in a new centrifuge tube (supplied), add 50-100 µl Buffer EB to the middle of the adsorbent membrane, leave it at room temperature for 2 minutes, centrifuge at 13,000 rpm for 1 minute, and collect the plasmid solution into the centrifuge tube. -The plasmid solution was collected into the centrifuge tube.Note: 1) To increase the recovery efficiency of the plasmid, the resulting solution can be reintroduced into the adsorbent column, left at room temperature for 2 minutes, centrifuged at 13,000 rpm for 1 minute, and the plasmid solution collected into a centrifuge tube.2) For low plasmid copy number or >10 kb, Buffer EB is preheated at 65-70°C in a water bath to increase extraction efficiency... Read More | Product DescriptionAcetyl esterase (sialate-O-acetylesterase) is a recombinant protein from Tannerella forsythia, ATCC 43037 strain, expressed in Escherichia coli. The enzyme removes acetyl groups attached via an O- group, mainly 9-, 8- and 7-. It can be used for monitoring of diacetylation of Product DescriptionAcetyl esterase (sialate-O-acetylesterase) is a recombinant protein from Tannerella forsythia, ATCC 43037 strain, expressed in Escherichia coli. The enzyme removes acetyl groups attached via an O- group, mainly 9-, 8- and 7-. It can be used for monitoring of diacetylation of sialic acids on products such as erythropoietin (EPO).The Zyme Acetyl Esterase Kit removes 9-, 8- and 7-O-acetyl groups from released sialic acids, released glycans or glycoproteins. It is commonly used for the characterization of highly-sialylated biotherapeutics such as EPO, FSH and blood clotting factors.Molecular Weight76.3 kDContentsAcetyl esterase – PBS pH7.5 buffer containing 10 mM Tris-HClReaction Buffer – 500 mM sodium acetate pH5.5Number of SamplesSufficient for up to 50 samples.Amount of SampleUp to 10 µg glycoprotein, up to 2.5 µg released glycans and up to 1 µg free sialic acid per digestion.Suitable SamplesAcetyl esterase (sialate-O-acetylesterase) can act upon complex glycoprotein samples, such as erythropoietin (EPO), bovine submaxillary mucin and oral epithelial cell-bound glycans, and on N- and O-glycans released from a glycoprotein. Either fluorescently labelled or unlabelled glycans are suitable. It can also be used on released sialic acids.Unit DefinitionOne unit (U) of acetyl esterase is defined as the amount of enzyme required to produce 300 µmole of 4-nitrophenol and acetate in 1 minute at 30°C in a buffer containing 50 mM Tris-HCl, 140 mM NaCl, pH 8.5, from 4-nitrophenyl acetate, a chromogenic esterase substrateStorageProtect from sources of heat and light. When stored correctly, the enzyme should be stable for 24 months from date of purchase. Exposure to ambient temperatures (20 – 26°C) over 3 days does not result in a reduction of enzymatic activity.ShippingThe product should be shipped at 4°C.HandlingEnsure that any glass, plastic ware or solvents used with this item are free of environmental carbohydrates. Use powder-free gloves for all sample handling procedures and avoid contamination with environmental carbohydrate.SafetyPlease read the Safety Data Sheets (SDSs) for all chemicals used. All processes involving labelling reagents should be performed using appropriate personal safety protection – safety glasses, chemically resistant gloves (e.g. nitrile), lab coat, and when appropriate, in a laboratory fume cupboard.For research use only. Not for human or drug use ApplicationAcetyl esterase (sialate-O-acetylesterase) can be used to remove 9-, 8- and 7-O-acetyl groups from released sialic acids, released glycans or glycoproteins... Read More |