本试剂盒适用于从700 μL 血浆、血清、尿液中提取游离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产物可直接用于荧光定量PCR、PCR、生物芯片分析、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。
| M1522674 | 组件名称
| 50T | 储存
|
| M1522674A | 蛋白酶K
| 1.1 mL | 2-8℃.
|
| M1522674B | 磁珠MXVB-CF
| 1.1 mL
| 2-8℃.
|
| M1522674C | 裂解结合液CLB
| 55 mL | RT.
|
| M1522674D | 洗涤液DW1
| 80 mL | RT.
|
| M1522674E | 洗涤液DW2
| 70 mL | RT.
|
| M1522674F | 洗脱液DE
| 1.8 mL | RT. |
储存条件
M1522674A, M1522674B：2-8℃保存，12个月。
M1522674C, M1522674D, M1522674E, M1522674F：室温保存，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，裂解液CLB出现沉淀，需要30℃水浴至溶液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：700 μL的血浆、血清、尿液样本。
- 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
3. 试剂准备
4. 操作步骤
磁分离操作建议（步骤4.2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 将样本平衡至室温，样本及各试剂涡旋混匀后，取2.0 mL离心管，依次加入1 mL游离核酸裂解结合液CLB、700 µL刚分离血浆或刚解冻血浆、20 µL蛋白酶K和20 µL磁珠MXVB-CF。涡旋混匀后室温裂解10分钟，期间每隔3分钟涡旋一次，每次10秒，然后将离心管放入振荡仪振荡或放到涡旋仪涡旋5分钟，使DNA尽量吸附到磁珠上。
备注：请避免将蛋白酶K与裂解结合液CLB预先混合。
4.2 转移至磁力架上进行磁分离至溶液透亮，吸弃溶液。
4.3加入700 µL洗涤液DW1，浸泡30秒，再涡旋10秒打散磁珠后转至磁力架上进行磁分离至溶液澄清，吸弃溶液。
4.4 再次加入700 µL洗涤液DW1，涡旋10秒打散磁珠后转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，涡旋10秒打散磁珠后转至磁力架上进行磁分离至溶液澄清，吸弃溶液。
4.6 再次加入500 µL洗涤液DW2，涡旋10秒打散磁珠后转至磁力架上进行磁分离至溶液澄清，吸弃溶液。
4.7 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。
4.8 加入30 µL洗脱液DE，高速涡旋1~3分钟打散磁珠。
4.9 60℃温浴5分钟，温浴期间取出涡旋3次，每次10秒，温浴结束后高速涡旋30秒。
4.10 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。
4.11 把游离DNA溶液转移至新的离心管中待用。如果不立即使用，请存储于-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 suitable for extracting free DNA from 700 µL of plasma, serum, or urine. It utilizes superparamagnetic particle purification technology, eliminating the need for toxic phenol‑chloroform extraction. The procedure is simple and convenient, and the obtained DNA product can be directly used in downstream experiments such as real‑time PCR, PCR, biochip analysis, first‑generation sequencing, and next‑generation sequencing. The processed product is intended for research use only.
M1522674
| Component | 50T
| Storage |
|---|
| M1522674A | Proteinase K | 1.1 mL | 2–8 °C | | M1522674B | Magnetic Beads MXVB‑CF | 1.1 mL | 2–8 °C | | M1522674C | Lysis/Binding Buffer CLB | 55 mL | RT | | M1522674D | Wash Buffer DW1 | 80 mL | RT | | M1522674E | Wash Buffer DW2 | 70 mL | RT | | M1522674F | Elution Buffer DE | 1.8 mL | RT |
|
Storage Conditions
M1522674A, M1522674B: Store at 2–8 °C for 12 months.
M1522674C, M1522674D, M1522674E, M1522674F: Store at room temperature (RT) for 12 months.
Compatible Instruments
Manual extraction
Precautions
Read this instruction manual carefully before use.
Several buffers in this kit contain irritating guanidine salts. Wear gloves and follow standard safety precautions. Do not allow buffers to contact skin, eyes, or mucous membranes. If contact occurs, rinse immediately with plenty of water and seek medical advice.
If precipitation is observed in Lysis/Binding Buffer CLB due to low temperature, warm in a 30 °C water bath until the solution is completely clear before use.
If magnetic beads cannot be resuspended by vigorous vortexing due to low temperature, do not use them.
Several buffers in this kit contain guanidine salts. Do not treat with oxidizing disinfectants such as sodium hypochlorite, as toxic gas may be released. Dispose as medical waste.
The components of this kit are intended to be used together. Do not interchange or mix components from different lot numbers.
Usage Method
1. Sample Requirements
Specimen type: 700 µL plasma, serum, or urine.
Specimen storage and transport: Specimens can be used immediately or stored at –70 °C or lower for up to 6 months; avoid repeated freeze‑thaw cycles. Transport under冷链 (cold chain) conditions.
Freeze‑thaw recommendation: Rapid freezing and thawing; avoid repeated freeze‑thaw cycles.
2. Materials Required but Not Provided
Constant temperature water bath (for dissolving precipitates)
Dry thermostat incubator
1.5/5.0 mL sterile, nuclease‑free centrifuge tubes
Various sizes of pipettes and tips (20, 200, 1000 µL)
3. Reagent Preparation
Check solutions for precipitates and verify that magnetic beads can be resuspended.
4. Procedure
Recommendation for magnetic separation (starting from step 4.2): After placing the centrifuge tube on the magnetic stand, gently rotate left and right. Once the magnetic beads have gathered on the tube wall near the magnet, gently invert the magnetic stand several times to collect beads on the cap wall as well. Let stand until the solution becomes clear (standing time depends on the magnet strength).
4.1 Equilibrate the sample to room temperature. Vortex the sample and all reagents. In a 2.0 mL centrifuge tube, sequentially add 1 mL Free DNA Lysis/Binding Buffer CLB, 700 µL freshly isolated or freshly thawed plasma, 20 µL Proteinase K, and 20 µL Magnetic Beads MXVB‑CF. Vortex to mix, then lyse at room temperature for 10 minutes, vortexing for 10 seconds every 3 minutes. Then place the tube on a shaker or vortex for 5 minutes to maximize DNA adsorption onto the beads.
Note: Do not pre‑mix Proteinase K with Lysis/Binding Buffer CLB.
4.2 Transfer the tube to a magnetic stand and perform magnetic separation until the solution is clear. Aspirate and discard the supernatant.
4.3 Add 700 µL Wash Buffer DW1, soak for 30 seconds, then vortex for 10 seconds to resuspend the beads. Place on the magnetic stand, separate until clear, and discard the supernatant.
4.4 Add another 700 µL Wash Buffer DW1, vortex for 10 seconds to resuspend the beads, then separate on the magnetic stand until clear and discard the supernatant.
4.5 Add 700 µL Wash Buffer DW2, vortex for 10 seconds to resuspend the beads, then separate on the magnetic stand until clear and discard the supernatant.
4.6 Add another 500 µL Wash Buffer DW2, vortex for 10 seconds to resuspend the beads, then separate on the magnetic stand until clear and discard the supernatant.
4.7 Briefly centrifuge in a microcentrifuge, then place on the magnetic stand until the solution is clear. Aspirate and discard all remaining liquid. Open the tube cap and air‑dry for 3–5 minutes.
Note: Residual ethanol will inhibit downstream enzymatic reactions, so ensure that the ethanol is completely evaporated during drying. Do not over‑dry, as this may affect subsequent elution efficiency.
4.8 Add 30 µL Elution Buffer DE, and vortex vigorously for 1–3 minutes to resuspend the beads.
4.9 Incubate at 60 °C for 5 minutes. During the incubation, remove the tube and vortex 3 times, 10 seconds each time. After incubation, vortex vigorously for 30 seconds.
4.10 Briefly centrifuge to collect droplets from the cap, then place on the magnetic stand for magnetic separation.
4.11 Transfer the free DNA solution to a new centrifuge tube for later use. If not used immediately, store at –15 to –25 °C; for long‑term storage, keep at –70 °C or lower.
Limitations of the Method
For some poorly collected samples, the extracted product may contain minor impurities that inhibit PCR amplification.
The efficiency of nucleic acid extraction is closely related to the quality of sample collection, processing, transport, and storage. Any errors in these steps may lead to inaccurate results.
I2611510
危险品化学品经营许可证（带存储）