| 货号 (SKU) | 包装规格 | 是否现货 | 价格 | 数量 |
|---|---|---|---|---|
| N667427-200T |
200T |
期货  |
| |
| N667427-50T |
50T |
期货 |
|
| 英文名称 | NuClean Plant Genomic DNA Kit | ||||||||||||||||||||||||||||||||||||||||||||||||||
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| 储存温度 | 室温 | ||||||||||||||||||||||||||||||||||||||||||||||||||
| 运输条件 | 常规运输 | ||||||||||||||||||||||||||||||||||||||||||||||||||
| 产品介绍 |
产品内容
产品简介 本试剂盒采用高效、专一结合核酸的离心吸附柱和独特的缓冲系统，适合从各种不同的新鲜或冻存植物组织中提取基因组DNA，并可最大限度去除植物组织中的杂质。本试剂盒无需使用酚/氯仿抽提，操作安全。提取的基因组DNA片段大、纯度高、质量稳定可靠， 适用于PCR、荧光定量PCR、分子标记、文库构建等实验。 自备试剂：无水乙醇 实验前准备及重要注意事项 1. 样品应避免反复冻融，否则会导致提取的DNA片段较小且提取量下降。 2.第一次使用前应按照试剂瓶标签的说明在Buffer LP3和Buffer GW2中加入无水乙醇。使用前请检查Buffer LP1和Buffer LP2是否出现结晶或者沉淀，如有结晶或者沉淀，请将Buffer LP1和Buffer LP2于56℃水浴重新溶解。 操作步骤 1. 取植物新鲜组织100 mg左右或干重组织约20 mg，加入液氮充分研磨。 2. 将研磨后的粉末收集到离心管（自备）中，加入400 μl Buffer LP1和6 μl RNase A（10 mg/ml），涡旋振荡1分钟，室温放置10分钟，使其充分裂解。 注意：1）使用涡旋振荡或移液器吹打，充分裂解组织，组织裂解不完全会影响最终的DNA得率。2）请勿在使用前将Buffer LP1与RNase A混合。 3. 加入130 μl Buffer LP2，混匀，涡旋震荡1分钟。 4.12,000 rpm（~13,400×g）离心5分钟，将上清移至新的离心管（自备）中。 5. 加入1.5倍体积的Buffer LP3（使用前检查是否已加入无水乙醇），充分混匀（如500 μl滤液加入750 μl Buffer LP3）。 注意：加入Buffer LP3后应立即混匀，可能会产生沉淀但不影响后续实验。 6.将上步所得溶液和沉淀全部加入到已装入收集管的吸附柱（Spin Columns DM）中，若一次不能加完溶液，可分多次转入。12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。 7. 向吸附柱中加入500 μl Buffer GW2（使用前检查是否已加入无水乙醇），12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。 注意：如吸附膜呈现绿色，向吸附柱中加入500 μl无水乙醇，12,000 rpm离心1分钟，倒掉收集管中的废液，将吸附柱重新放回收集管中。 8. 重复步骤7. 9．12,000 rpm离心2分钟，倒掉收集管中的废液。将吸附柱置于室温数分钟，以彻底晾干。注意：这一步的目的是将吸附柱中残余的乙醇去除，乙醇的残留会影响后续的酶促反应（酶切、PCR 等）。 10.将吸附柱放到一个新离心管（自备）中，向吸附膜的中间部位悬空滴加50-100 μl Buffer GE或灭菌水，室温放置2-5分钟，12,000 rpm离心1分钟，收集DNA溶液。-20℃ 保存DNA。 注意：1）如果下游实验对pH值或EDTA敏感，可以用灭菌水洗脱。洗脱液的pH值对洗脱效率有很大影响，若用水做洗脱液应保证其pH值在7.0-8.5（可以用NaOH将水的pH值调到此范围），pH 值低于7.0时洗脱效率不高。 2）离心之前室温孵育5分钟可以增加产量。 3）如果要提高DNA的终浓度，可以将步骤10所得的DNA洗脱液重新加至吸附膜上，重复步骤10； 若洗脱体积小于100 μl，可以增加DNA的终浓度，但可能会减少DNA的总产量。如果所得DNA的量 小 于 1µg， 推 荐 用 50μl Buffer GE 进行洗脱 。 4）因为保存在水中的DNA会受到酸性水解作用的影响，如需长期保存，推荐用Buffer GE洗脱并于-20℃保存。 Product content
Products This kit uses centrifugal adsorption columns with high efficiency and specific binding of nucleic acids and a unique buffer system, which is suitable for extracting genomic DNA from a wide variety of different fresh or frozen plant tissues with maximum removal of impurities from the plant tissues. The kit eliminates the need for phenol/chloroform extraction and is safe to handle. The extracted genomic DNA fragments are large, high purity, stable and reliable quality, suitable for PCR, fluorescence quantitative PCR, molecular labeling, library construction and other experiments. Self-contained reagent: anhydrous ethanol Pre-experiment Preparation and Important Notes 1. Repeated freezing and thawing of the sample should be avoided, as this may result in smaller fragments of extracted DNA and a decrease in the amount extracted. 2. Anhydrous ethanol should be added to Buffer LP3 and Buffer GW2 according to the instructions on the label of the reagent bottle before first use. Check Buffer LP1 and Buffer LP2 for crystallization or precipitation before use. If crystallization or precipitation occurs, re-dissolve Buffer LP1 and Buffer LP2 in a 56°C water bath. Procedure 1. Take about 100mg of fresh plant tissue or about 20mg of dry weight tissue and add liquid nitrogen to grind it fully. 2. Collect the ground powder into a centrifuge tube (self-provided), add 400 μl Buffer LP1 and 6 μl RNase A (10 mg/ml), vortex and oscillate for 1 minute, and leave it at room temperature for 10 minutes to allow for full cleavage. Note: 1) Use vortex shaking or pipette blowing to fully lyses the tissue, incomplete tissue lysis will affect the final DNA yield. 2) Do not mix Buffer LP1 with RNase A prior to use. 3. Add 130 μl Buffer LP2, mix well and vortex for 1 minute. 4. Centrifuge at 12,000 rpm (~13,400 x g) for 5 minutes and transfer the supernatant to a new centrifuge tube (supplied). 5. Add 1.5 times the volume of Buffer LP3 (check that anhydrous ethanol has been added before use) and mix thoroughly (e.g., 500 μl filtrate to 750 μl Buffer LP3). Note: Buffer LP3 should be mixed immediately after addition; precipitation may occur but will not affect subsequent experiments. 6. Add all of the solution and precipitate obtained in the previous step to the adsorption columns (Spin Columns DM) that have been loaded into the collection tubes, if the solution cannot be added all at once, it can be transferred in several times. centrifuge the columns at 12,000 rpm for 1 minute, pour off the waste liquid in the collection tubes, and put the columns back into the collection tubes. 7. Add 500 μl of Buffer GW2 to the adsorption column (check that anhydrous ethanol has been added before use), centrifuge at 12,000 rpm for 1 minute, pour off the waste liquid in the collection tube, and put the adsorption column back into the collection tube. Note: If the adsorbent membrane appears green, add 500 μl of anhydrous ethanol to the adsorbent column, centrifuge the column at 12,000 rpm for 1 minute, pour off the waste liquid in the collection tube, and put the adsorbent column back into the collection tube. 8. Repeat step 7. 9. Centrifuge at 12,000 rpm for 2 minutes and pour off the waste liquid in the collection tube. Leave the adsorption column at room temperature for several minutes to dry thoroughly. Note: The purpose of this step is to remove residual ethanol from the adsorption column, which can interfere with subsequent enzymatic reactions (digestion, PCR, etc.). 10. Place the adsorption column in a new centrifuge tube (supplied), add 50-100 μl of Buffer GE or sterilized water dropwise to the middle of the adsorbent membrane, leave it at room temperature for 2-5 minutes, and centrifuge it at 12,000 rpm for 1 minute to collect the DNA solution. -The DNA solution was collected by centrifugation at 12,000 rpm for 1 min. Note: 1) If the downstream experiment is sensitive to pH or EDTA, you can use sterilized water for elution. The pH value of the eluent has a great influence on the elution efficiency, if you use water as the eluent, you should ensure that the pH value is 7.0-8.5 (you can use NaOH to adjust the pH value of the water to this range), and when the pH value is lower than 7.0, the elution efficiency is not high. 2) Incubation at room temperature for 5 minutes prior to centrifugation increases yield. (3) If the final concentration of DNA is to be increased, the DNA eluate obtained in step 10 can be re-added to the adsorbent membrane and repeat step 10; if the elution volume is less than 100µl, the final concentration of DNA can be increased, but it may reduce the total DNA yield. If the amount of DNA obtained is less than 1µg, 50µl Buffer GE is recommended for elution. 4) Because DNA stored in water is subject to acidic hydrolysis, for long-term storage, elution with Buffer GE and storage at -20°C are recommended. |
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