| 货号 (SKU) | 包装规格 | 是否现货 | 价格 | 数量 |
|---|---|---|---|---|
| S666146-50T |
50T |
期货  |
| |
| S666146-200T |
200T |
期货 |
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| 英文名称 | Swab 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 本试剂盒提供一种简单快速分离纯化口腔拭子样本总DNA的方法。该试剂盒采用可特异性结合DNA的硅基质膜和独特的缓冲系统，高效专一吸附DNA，每个拭子可得到0.5-3.5 μg基因组DNA，提取的DNA片段大、纯度高、质量稳定可靠。适用于酶切、PCR、文库构建、Southern杂交等实验。 自备试剂：无水乙醇。 实验前准备及重要注意事项 1. 第一次使用前应按照试剂瓶标签说明在Buffer GW1和Buffer GW2中加入无水乙醇。 2. 使用前若发现Buffer GL有沉淀，请将Buffer GL于56℃水浴溶解。 3. 全部离心步骤可在室温下进行。 4. 取样：使用口腔拭子在口腔内壁擦拭6次，晾干2小时保存，为确保样本不被食物或 饮料污染，取样前30分钟内请勿进食和饮水。 操作步骤 1. 将口腔拭子的棉签用剪刀从杆上剪下，置于2 mL的离心管（自备）中，加入400 μL Buffer GR。 注意：如需无RNA污染的基因组DNA，可加入4 μL浓度为100 mg/ml 的RNase A溶液震荡混匀。 2. 加入20 μL Proteinase K 和 400 μL Buffer GL，立即涡旋震荡15秒，充分混匀。 注意：加入Buffer GL后立即充分混匀；不可将Proteinase K直接加入Buffer GL中使用。 3．56℃放置10分钟，短暂离心，使管壁上的溶液收集到管底。 4. 加入400 μL无水乙醇，涡旋震荡充分混匀，短暂离心，使管壁上的溶液收集到管底。 注意：加入无水乙醇后可能会产生白色沉淀，不会影响后续实验。 5. 将上步所得溶液和沉淀分两次加入到已装入收集管的吸附 柱（Spin Columns DS） 中，一次最多不超过700 μL 。12,000 rpm（～13,400×g）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。 6. 向吸附柱中加入500 μL Buffer GW1（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。 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3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：如需进一步提高DNA纯度，可重复步骤7。 8．12,000 rpm离心1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）。 9．将吸附柱置于一个新的1.5 mL离心管中，向吸附柱的中间部位悬空加入50 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。 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）若需长期保存，推荐用Buffer GE洗脱并于-20℃保存。
Product content
Products This kit provides a simple and rapid method for the isolation and purification of total DNA from buccal swab samples. The kit adopts a silica matrix membrane that can specifically bind DNA and a unique buffer system to adsorb DNA efficiently and specifically, and 0.5-3.5 μg of genomic DNA can be obtained from each swab, and the extracted DNA fragments are large, pure and of stable and reliable quality. It is suitable for enzyme digestion, PCR, library construction, Southern hybridization and other experiments. Self-contained reagent: anhydrous ethanol. Pre-experiment Preparation and Important Notes 1. Anhydrous ethanol should be added to Buffer GW1 and Buffer GW2 according to the instructions on the label of the reagent bottle before first use. 2. If precipitation is found in Buffer GL before use, dissolve Buffer GL in a 56°C water bath. 3. All centrifugation steps can be performed at room temperature. 4. Sampling: Use a buccal swab to wipe the inside of the mouth 6 times, dry for 2 hours and store. To ensure that the sample is not contaminated by food or drink, do not eat or drink for 30 minutes before sampling. Procedure 1. The swab of the buccal swab was cut from the rod with scissors and placed in a 2mL centrifuge tube (supplied) and 400μL Buffer GR was added. Note: For genomic DNA without RNA contamination, add 4 μL of RNase A solution at a concentration of 100 mg/ml and shake to mix. 2. Add 20 μL of Proteinase K and 400 μL of Buffer GL, immediately vortex and shake for 15 seconds and mix thoroughly. Note: Mix well immediately after adding Buffer GL; do not add Proteinase K directly to Buffer GL for use. 3.56°C for 10 minutes and centrifuge briefly so that the solution on the walls of the tube collects at the bottom. 4. Add 400 μL of anhydrous ethanol, vortex and shake to mix thoroughly, and centrifuge briefly so that the solution on the wall of the tube collects at the bottom of the tube. Note: The addition of anhydrous ethanol may produce a white precipitate that will not affect subsequent experiments. 5. Add the solution and precipitate obtained in the previous step to the Spin Columns DS in two batches of up to 700 μL at a time into the collection tube. centrifuge the column at 12,000 rpm (∼13,400 × g) for 1 minute, pour off the waste liquid from the collection tube, and return the column to the collection tube. 6. Add 500 μL of Buffer GW1 to the adsorbent 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 adsorbent column back into the collection tube. 7. Add 500 μL of Buffer GW2 to the adsorption column (check that anhydrous ethanol has been added before use), centrifuge the column at 12,000 rpm for 3 minutes, pour off the waste liquid in the collection tube, and put the column back into the collection tube. Note: Step 7 can be repeated if further DNA purity is required. 8. Centrifuge at 12,000 rpm for 1 minute 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.). 9. Place the adsorption column in a new 1.5 mL centrifuge tube, add 50 μL of Buffer GE or sterilized water to the middle of the adsorption column overhanging the column, let stand at room temperature for 2-5 minutes, centrifuge at 12,000 rpm for 1 minute, collect the DNA solution, and store at -20℃. Attention: (1) If the downstream experiment is sensitive to pH or EDTA, it can be eluted with sterilized water. The pH value of the eluent has a great influence on the elution efficiency. If the eluent is made of water, the pH value should be 7.0-8.5 (the pH value of water can be adjusted to this range by using NaOH), and the elution efficiency is not high when the pH value is lower than 7.0. 2) For long-term storage, it is recommended to elute with Buffer GE and store at -20°C. |
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