本试剂盒适用于从200 μL PCR产物、洗脱产物或酶促反应产物中回收高纯度的DNA，该方法采用超顺磁性的磁性粒子纯化技术，提取过程中无需使用有毒的酚氯仿抽提，操作简单方便，得到的DNA产物可直接用于PCR、荧光定量PCR、芯片分析、一代、二代测序等相关实验。其处理后的产物用于科研检测使用。
储存条件
M1521715A, M1521715C, M1521715D：室温保存，12个月。
M1521715B：2-8℃保存，12个月。
适配仪器
手工提取
注意事项
1. 使用前请仔细阅读本说明书。
2. 本试剂盒中的多种缓冲液含有刺激性的胍盐，务必戴上手套，并按照安全标准预防措施来处理。不要让缓冲液接触到皮肤、眼睛以及黏膜，如果确实发生，请立即用大量清水清洗并就医。
3. 如因气温较低，溶液出现沉淀，需要30℃水浴至沉淀完全溶解后方可使用。
4. 如因气温较低，用力振荡后磁珠无法重悬，请勿使用。
5. 本试剂盒中的多种缓冲液含胍盐，请勿用氧化性消毒剂如次氯酸钠进行处理，否则会释放有毒气体，须按医疗废物进行处理。
6. 本试剂盒需配套使用，不同批号试剂盒中各组分不可以互换或混用。
使用方法
1. 样本要求
- 适用标本类型：200 μL PCR产物、洗脱产物或酶促反应产物。
- 标本保存和运输：标本可立即用于测试，也可以保存于-70℃或更低温度待测，保存期为6个月，应避免反复冻融。标本运输采用冷链运输。
- 冻融要求：速冻速融，避免反复冻融。
2. 自备试验器材
恒温水浴锅（当溶液出现沉淀时使用）
磁力架（1.5/2.0 mL）、1.5/2.0 mL无菌无核酸酶离心管、各种规格移液器及Tips（20、200、1000 μL）
3. 试剂准备
检查溶液是否有沉淀，磁珠是否能重悬。
4. 操作步骤
磁分离操作建议（步骤4.4开始）：离心管置于磁力架后，轻轻地左右旋转，待磁珠聚集于贴近磁力架的管壁后，轻轻地颠倒磁力架数次，使管盖上的磁珠也聚集到管壁，静置数分钟至溶液澄清即可（静置时长视磁力架磁性而定）。
4.1 取200 µL样本于1.5 ml离心管中，若样本体积小，用PBS或无菌无核酸酶水补至200 µL。
4.2 加入600 µL结合液CP，涡旋混匀。
4.3 加入20 µL磁珠MXVB，振荡或涡旋5~7分钟使磁珠吸附DNA。
4.4 转移至磁力架上进行磁分离至溶液澄清，吸弃溶液。
4.5 加入700 µL洗涤液DW2，取出涡旋10秒打散磁珠后放至磁力架进行磁分离至溶液澄清，吸弃溶液。
4.6 掌上离心机瞬时离心后，转至磁力架上吸附至溶液澄清，充分吸弃所有溶液，打开管盖，空气干燥3~5分钟。
注意：乙醇残留会抑制后续的酶反应，所以晾干时要确保乙醇挥发干净。切勿干燥过久，以免影响后续洗脱效果。
4.7 加入20~100 µL洗脱液DE，高速涡旋2~3分钟打散磁珠。
4.8 65℃温浴6分钟，然后高速涡旋30秒。
4.9 瞬时离心收集管盖液滴至管中，转移至磁力架上进行磁分离。
4.10 把DNA溶液转移至新的离心管中，如果不立即使用，存储于-15~-25℃，长期保存请放置于-70℃或更低的温度。
方法的局限性
1. 对于某些取样不佳的样本，提取产物中可能存在少量杂质抑制PCR扩增。
2. 样本核酸的提取效果与样本收集、处理、运输和保存质量密切相关，其中任何失误都将会导致结果不准确。
This kit is designed for the recovery of high-purity DNA from 200 μL of PCR products, elution products, or enzymatic reaction products. The method employs superparamagnetic particle purification technology. The extraction process eliminates the need for toxic phenol-chloroform extraction. The procedure is simple and convenient, and the resulting DNA product can be used directly in downstream applications such as PCR, quantitative real-time PCR, microarray analysis, and first- and second-generation sequencing. The processed products are intended for scientific research and testing purposes only.
Storage Conditions
M1521715A, M1521715C, M1521715D: Store at room temperature, stable for 12 months.
M1521715B: Store at 2–8°C, stable for 12 months.
Compatible Instruments
Manual extraction.
Precautions
Read this manual carefully before use.
Several buffers in this kit contain irritating guanidinium salts. Always wear gloves and handle according to standard safety precautions. Avoid contact of buffers with skin, eyes, and mucous membranes. In case of contact, flush immediately with plenty of water and seek medical attention.
If precipitation occurs in the solution due to low ambient temperatures, incubate at 30°C in a water bath until the precipitate is completely dissolved before use.
If the magnetic beads cannot be resuspended after vigorous shaking due to low ambient temperatures, do not use the beads.
Several buffers in this kit contain guanidinium salts. Do not treat with oxidizing disinfectants such as sodium hypochlorite, as this may release toxic gases. Dispose of waste according to medical waste regulations.
This kit components are designed to be used together as a system. Components from different lot numbers should not be interchanged or mixed.
Instructions for Use
1. Sample Requirements
Applicable sample type: 200 μL PCR product, elution product, or enzymatic reaction product.
Sample storage and transport: Samples may be processed immediately or stored at -70°C or lower for up to 6 months. Avoid repeated freeze-thaw cycles. Transport samples using cold chain logistics.
Freeze-thaw requirements: Rapid freezing and rapid thawing are recommended; avoid repeated freeze-thaw cycles.
2. Equipment and Materials to be Supplied by User
Constant temperature water bath (for use when precipitation occurs in solutions)
Magnetic separation rack (for 1.5/2.0 mL tubes)
1.5/2.0 mL sterile, nuclease-free centrifuge tubes
Pipettes and tips of various volumes (20, 200, 1000 μL)
3. Reagent Preparation
Check solutions for any precipitation and confirm that the magnetic beads can be resuspended.
4. Operating Procedure
Recommendation for magnetic separation (starting from step 4.4): After placing the centrifuge tube on the magnetic rack, gently rotate it left and right. Once the magnetic beads have aggregated against the tube wall closest to the magnet, gently invert the entire rack several times to collect any beads from the cap onto the tube wall. Allow to stand for several minutes until the solution becomes clear (standing time depends on the strength of the magnetic rack).
4.1 Transfer 200 μL of sample to a 1.5 mL centrifuge tube. If the sample volume is less than 200 μL, adjust the volume to 200 μL with PBS or sterile, nuclease-free water.
4.2 Add 600 μL of Binding Buffer CP and mix thoroughly by vortexing.
4.3 Add 20 μL of Magnetic Beads MXVB and shake or vortex for 5–7 minutes to allow DNA binding to the beads.
4.4 Place the tube on the magnetic rack for magnetic separation until the solution is clear, then aspirate and discard the supernatant.
4.5 Add 700 μL of Wash Buffer DW2. Remove the tube from the rack, vortex for 10 seconds to disperse the beads, then place back on the magnetic rack for separation until the solution is clear. Aspirate and discard the supernatant.
4.6 Perform a quick spin using a mini centrifuge, then place the tube back on the magnetic rack and allow beads to collect until the solution is clear. Carefully aspirate and discard all residual liquid. Open the tube cap and air-dry the beads for 3–5 minutes.
Note: Residual ethanol will inhibit downstream enzymatic reactions. Ensure ethanol has completely evaporated during drying. Do not over-dry the beads, as this may affect subsequent elution efficiency.
4.7 Add 20–100 μL of Elution Buffer DE and vortex at high speed for 2–3 minutes to fully disperse the beads.
4.8 Incubate at 65°C for 6 minutes, then vortex at high speed for 30 seconds.
4.9 Briefly centrifuge to collect any droplets from the cap into the tube, then place on the magnetic rack for magnetic separation.
4.10 Transfer the DNA solution to a new centrifuge tube for subsequent use. If not used immediately, store at -15 to -25°C. For long-term storage, place at -70°C or lower.
Limitations of the Method
For certain poorly collected samples, trace impurities in the extracted product may inhibit PCR amplification.
The efficiency of nucleic acid extraction is closely related to the quality of sample collection, handling, transport, and storage. Any error in these processes may lead to inaccurate results.
G2618004
危险品化学品经营许可证（带存储）