本试剂盒适用于深加工食品DNA的抽提。深加工食品经过煮、煎、炸、腌制和发酵等数道工序制备而成，原材料经过高温、机械损伤和极端环境处理后，基因组DNA已严重断裂和降解，故我司采用了超顺磁性的磁性粒子纯化技术和CTAB裂解法，保证深加工食品可以充分裂解，释放出其中的DNA，再通过一系列的处理将其中的杂质清除，得到的上清液采用磁珠吸附后，经过洗涤清除杂质，最后在最佳的洗脱条件下获得DNA。本试剂盒纯化的DNA可直接用于后续PCR、QPCR检测。其处理后的产物用于科研检测使用。
| M1522683 | 组件 | 物理外观 | 48T | 储存 | | M1522683A | 蛋白酶K | 液体 | 1.1 mL | 2-8℃. | | M1522683B | 裂解液CTAB | 液体 | 40 mL | RT. | | M1522683C | 分层液SF | 液体 | 30 mL | 2-8℃. Store in the dark. | | M1522683D | 磁珠MXVB | 液体 | 1.1 mL | 2-8℃. | | M1522683E | 洗涤液PW1 | 液体 | 40 mL | RT. | | M1522683F | 洗涤液BW1 | 液体 | 40 mL | RT. | | M1522683G | 洗涤液DW2 | 液体 | 40 mL | RT. | | M1522683H | 洗脱液DE | 液体 | 6 mL | RT. |
|
储存条件
M1522683A, M1522683D：2-8℃保存，12个月。
M1522683C：2-8℃避光保存，12个月。
M1522683B, M1522683E, M1522683F, M1522683G, M1522683H：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，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，裂解液CTAB出现沉淀，可60℃水浴至沉淀完全溶解后使用。
5.如因气温较低，用力振荡后磁珠无法重悬，请勿使用。
6. 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。
7. 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：深加工食品（如意大利面、方便面、薯片、腐乳、腐竹等）。
- 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。
2. 自备试验器材
3. 试剂准备
4. 操作步骤
4.1 样本处理
- 固态食品：取足量的固态食品如意大利面、方便面、薯片、腐乳、腐竹等，液氮或者机械研磨至粉末，称取100~200 mg样本至2.0 mL离心管中。
- 豆浆等液态食品：取20 mL液态食品于50 mL离心管中，12,000×g离心10分钟，弃上清。
- 加工处理的干燥植物组织及中草药：称取100~300 mg经液氮研磨的干燥的植物组织或者中草药粉末至2.0 mL离心管中。
4.2 加入700 μL裂解液CTAB和20 µL蛋白酶K，涡旋混匀样本，使样本与裂解液充分混合（若样本较干燥，可在离心管中预先加入700 μL裂解液CTAB再加入样本，利于样本与裂解液混合）。
注意：依据食品材料的干燥度、体积和大小，可以适量调节裂解液CTAB的用量，食品材料需要完全浸没在裂解液CTAB中才能保证充分裂解。
4.3 56℃振荡温浴60分钟，如恒温仪没有振荡功能，温浴期间可每15分钟涡旋30秒促进裂解。
4.4 取出样本冷却到室温，缓慢加入550 μL分层液SF，振荡充分混匀。
4.5 13,000×g离心5分钟。
4.6 吸取500 µL离心后的上层水相溶液，加入500 µL无水乙醇，高速涡旋混匀10秒。
4.7 加入20 µL磁珠MXVB，振荡或涡旋5~7分钟使磁珠吸附DNA。
4.8 转移至磁力架上进行磁分离至溶液澄清，吸弃溶液。
4.9 加入700 µL洗涤液PW1，涡旋30秒打散磁珠后转至磁力架进行磁分离至溶液澄清，吸弃溶液。
4.10 加入700 µL洗涤液BW1，涡旋30秒打散磁珠后转至磁力架进行磁分离至溶液澄清，吸弃溶液。
4.11 加入700 µL洗涤液DW2，涡旋30秒打散磁珠后转至磁力架进行磁分离至溶液澄清，吸弃溶液。
4.12 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。
4.13 加入30~60 µL洗脱液DE，高速涡旋2~3分钟打散磁珠。
4.14 60℃温浴5分钟，然后高速涡旋30秒。
4.15 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。
4.16 把基因组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 intended for DNA extraction from highly processed foods. Processed foods are prepared through multiple steps such as boiling, frying, deep‑frying, pickling, and fermentation. The raw materials undergo high temperature, mechanical damage, and extreme environments, resulting in severe fragmentation and degradation of genomic DNA. Therefore, our kit employs superparamagnetic particle purification technology combined with the CTAB lysis method to ensure thorough lysis of processed food samples, releasing their DNA. Impurities are removed through a series of treatments. The resulting supernatant is subjected to magnetic bead binding, followed by washing to remove impurities, and finally DNA is eluted under optimal conditions. DNA purified with this kit can be used directly for subsequent PCR and qPCR assays. The purified DNA is intended for research use only.
| M1522683 | Components | Appearance | 48T | Storage | | M1522683A | Proteinase K | Liquid | 1.1 mL | 2–8 °C | | M1522683B | Lysis Buffer CTAB | Liquid | 40 mL | RT | | M1522683C | Separation Solution SF | Liquid | 30 mL | 2–8 °C, Store in the dark. | | M1522683D | Magnetic Beads MXVB | Liquid | 1.1 mL | 2–8 °C | | M1522683E | Wash Buffer PW1 | Liquid | 40 mL | RT | | M1522683F | Wash Buffer BW1 | Liquid | 40 mL | RT | | M1522683G | Wash Buffer DW2 | Liquid | 40 mL | RT | | M1522683H | Elution Buffer DE | Liquid | 6 mL | RT |
|
Storage Conditions
M1522683A, M1522683D: Store at 2–8 °C for 12 months.
M1522683C: Store at 2–8 °C, protect from light, for 12 months.
M1522683B, M1522683E, M1522683F, M1522683G, M1522683H: Store at room temperature for 12 months.
Compatible Instrumentation
Manual extraction.
Precautions
Read the entire manual carefully before use.
Several buffers contain irritating guanidine salts. Always wear gloves and follow standard safety precautions. Avoid contact of buffers with skin, eyes, and mucous membranes. If contact occurs, rinse immediately with plenty of water and seek medical attention.
If precipitates appear due to low temperature, warm the solution at 30 °C until precipitates are completely dissolved before use.
If precipitates appear in Lysis Buffer CTAB due to low temperature, warm at 60 °C until completely dissolved before use.
If magnetic beads cannot be resuspended by vigorous shaking due to low temperature, do not use.
Several buffers contain guanidine salts. Do not treat with oxidizing disinfectants (e.g., sodium hypochlorite) as this may release toxic gas. Dispose as medical waste.
This kit is intended for use as an integrated system. Do not interchange or mix components from different lot numbers.
Protocol
1. Sample Requirements
Applicable sample types: Highly processed foods (e.g., pasta, instant noodles, potato chips, fermented tofu, dried bean curd sticks, etc.).
Sample preservation and transport: Samples can be processed immediately or stored at –70 °C or lower for up to 6 months. Avoid repeated freeze‑thaw cycles. Transport samples under cold chain conditions.
2. Materials to Be Prepared by User
Water bath (for dissolving precipitates), dry thermostat bath, liquid nitrogen, mortar and pestle, centrifuge
Magnetic stand (for 1.5/2.0 mL tubes)
1.5/2.0 mL sterile, nuclease‑free centrifuge tubes
Various pipettes and tips (20 µL, 200 µL, 1000 µL)
3. Reagent Preparation
Check solutions for precipitates and ensure magnetic beads can be resuspended.
4. Procedure
4.1 Sample Preparation
Solid foods: Take a sufficient amount of solid food (e.g., pasta, instant noodles, potato chips, fermented tofu, dried bean curd sticks). Grind to a fine powder using liquid nitrogen or a mechanical grinder. Weigh 100–200 mg of powder into a 2.0 mL centrifuge tube.
Liquid foods such as soy milk: Transfer 20 mL of liquid food into a 50 mL centrifuge tube. Centrifuge at 12,000×g for 10 minutes and discard the supernatant.
Processed dry plant materials and herbal medicines: Weigh 100–300 mg of dry plant material or herbal medicine powder (ground with liquid nitrogen) into a 2.0 mL centrifuge tube.
4.2 Add 700 µL of Lysis Buffer CTAB and 20 µL of Proteinase K. Vortex to mix the sample thoroughly (if the sample is very dry, add 700 µL of Lysis Buffer CTAB to the tube first, then add the sample – this facilitates mixing).
Note: Depending on the dryness, volume, and particle size of the food material, the amount of Lysis Buffer CTAB can be adjusted appropriately. The food material must be completely submerged in the lysis buffer to ensure efficient lysis.
4.3 Incubate at 56 °C with shaking for 60 minutes. If the incubator does not have a shaking function, vortex for 30 seconds every 15 minutes during incubation to promote lysis.
4.4 Cool the sample to room temperature. Slowly add 550 µL of Separation Solution SF and vortex vigorously to mix thoroughly.
4.5 Centrifuge at 13,000×g for 5 minutes.
4.6 Transfer 500 µL of the upper aqueous phase to a new tube. Add 500 µL of absolute ethanol and vortex vigorously for 10 seconds.
4.7 Add 20 µL of Magnetic Beads MXVB. Vortex or shake for 5–7 minutes to allow the beads to bind DNA.
4.8 Place the tube on a magnetic stand for magnetic separation until the solution is clear. Aspirate and discard the supernatant.
4.9 Add 700 µL of Wash Buffer PW1. Vortex for 30 seconds to resuspend the beads. Return to the magnetic stand and separate until the solution is clear. Aspirate and discard the supernatant.
4.10 Add 700 µL of Wash Buffer BW1. Vortex for 30 seconds to resuspend the beads. Separate magnetically until the solution is clear. Aspirate and discard the supernatant.
4.11 Add 700 µL of Wash Buffer DW2. Vortex for 30 seconds to resuspend the beads. Separate magnetically until the solution is clear. Aspirate and discard the supernatant.
4.12 Briefly centrifuge to collect residual liquid. Place the tube on the magnetic stand until the solution is clear. Aspirate and discard all remaining liquid thoroughly. Open the tube cap and air‑dry for 3–5 minutes.
Note: Residual ethanol will inhibit downstream enzymatic reactions. Ensure that ethanol is completely evaporated during drying. Do not over‑dry, as this may affect elution efficiency.
4.13 Add 30–60 µL of Elution Buffer DE. Vortex vigorously for 2–3 minutes to fully resuspend the beads.
4.14 Incubate at 60 °C for 5 minutes, then vortex vigorously for 30 seconds.
4.15 Briefly centrifuge to collect droplets from the lid. Return the tube to the magnetic stand for magnetic separation.
4.16 Transfer the genomic DNA solution to a new centrifuge tube for immediate 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 certain poorly collected samples, the extracted product may contain trace impurities that inhibit PCR amplification.
The efficiency of nucleic acid extraction is closely related to sample collection, processing, transport, and storage. Any error in these steps may lead to inaccurate results.
ZJ26F0636697
危险品化学品经营许可证（带存储）