Instrument Plus | Instrument Introduction: Nucleic Acid Extraction and Purification _ Operation Full-time Job
Nov 11th, 2022 at 00:37 Engineering Salford 248 viewsJob Details
if it is determined that the extraction effect is not good due to insufficient amount of magnetic beads, it is possible to improve the extraction effect by increasing the dosage of magnetic beads within a certain range. Taking GNT-02 series magnetic beads as an example, when extracting constant samples (plant tissues, whole blood, etc.),cbd crystallization equipment, the dosage is usually 10 UL/time; when extracting trace samples (such as serum free DNA, oral swabs, etc.), the dosage of magnetic beads is 15 ~ 20 UL/time. If you need to exceed this usage, you need to communicate with the technical engineer. 2. Mistake two The more reagents are used, the better the extraction effect is. The cracking effect is not good? Add more lysis solution. Washing effect is not good? Add more washing up liquid. This is the inertial thinking of many customers in the use of kits. However, for the magnetic bead method, every increase in the volume of a part of the liquid reduces the collision probability of more magnetic beads, and reducing the collision probability of magnetic beads will lead to a significant decline in the adsorption rate. Therefore, in many cases, although increasing the amount of lysis solution and washing solution can indeed play a role in enhancing lysis and washing, the core of magnetic bead extraction is the efficiency of magnetic bead adsorption of nucleic acid, which can not guarantee the efficiency of nucleic acid extraction, so simply increasing the amount of reagents used to improve the extraction effect may not be completely effective. For GNT-B02 whole blood genomic DNA kit, the amount of lysis solution should not exceed 400ul/time, and the amount of washing solution should not exceed 500ul/time. If an amplification system is required, the amount of magnetic beads and sample should be increased accordingly, and this amplification is not necessarily proportional. 3. Mistake three The more times of washing, the better the extraction effect. When there are too many impurities in the extracted nucleic acid, users will consider washing several times to obtain more pure nucleic acid. Increasing the number of times of washing is indeed conducive to purifying nucleic acid, but considering that each washing will lose a certain amount of nucleic acid and increase the possibility of nucleic acid cleavage and hydrolysis, it is generally appropriate to control the number of times of washing at 2 to 4 times. For GNT series kit, the washing times of single purification kit is 2 times, the washing times of plant and animal kit is 3 times, and the washing time of blood kit is 3-4 times. 4. Mistake four : The more samples taken, the better the extraction When the sample is not fresh enough or the nucleic acid content itself is very small, the nucleic acid extraction effect is often not good, many teachers will use the way of taking more samples to increase the amount of nucleic acid extraction. However, simply increasing the sample volume may sometimes introduce too many impurities, which will exceed the cracking capacity of the cracking solution and reduce the extraction efficiency. Therefore, it is not recommended to simply increase the sample volume to achieve the purpose of increasing the extraction volume. If the extraction volume is too low due to insufficient sample size, it is recommended to start the extraction after enrichment or concentration in the pretreatment. Or increasing the completeness of the cleavage to expose more nucleic acids is also a solution. 5. Mistake five If a certain kind of magnetic bead is good, it should work well in all tests. There are many kinds of magnetic beads, such as different particle size, different dispersion, different magnetic response time, different coating base matrix, different outer modified functional groups, different coating density and different arm length of functional groups, which will lead to different characteristics of magnetic beads. Therefore, different magnetic beads are suitable for different experiments and systems. Just like the same reagent for nucleic acid extraction, the formula is not exactly the same, cbd centrifugal extractor ,wiped film evaporator, and the properties of magnetic beads used for nucleic acid extraction are not exactly the same. Some magnetic beads show higher adsorption efficiency in the extraction of normal nucleic acid, and some magnetic beads are more suitable for the extraction of trace nucleic acid. if other media, websites or individuals reprint and use from this public account, please apply to the original author, and bear the copyright and other legal responsibilities. Instrument plus | dry goods finishing: chromatographic column use,wiped film distillation, maintenance and repair, all for you to organize together ~ do not collect it? Instrument Plus | News and Information: Southern Hemisphere Aerosol Content Breaks Record, Air Pollution Needs Urgent Attention! Instrument Plus | Dry Goods Finishing: Liquid Chromatography Basic Concepts and Terminology! Return to Sohu to see more Responsible Editor:. toptiontech.com