产品简介:
本产品综合MP公司的土壤DNA提取试剂盒产量高和Mobio公司纯度高的优点研制而成,采取MP公司完全一致的研磨管(3种研磨珠混合)方案可以处理更大的样品,可以直接使用MP研磨管一样的珠磨仪器和参数。独有的去腐殖酸技术可以完美去除各种腐殖酸和杂质的影响,提取的DNA纯度高,可以直接用于下游PCR反应。
操作步骤:
Before the first use, add the indicated amount of ethanol into PQ solution bottle, Buffer WB bottle, mix well, and mark the bottle with a check.
1:Add up to 500 mg of soil sample to a Bead Tube.
2:Add 980 μl Sodium Phosphate Buffer to sample in Bead Tube.Gentle votex to mix. Add 120 μl MT Buffer.
Note: Check MT Buffer. If MT Buffer is precipitated, heat solution to 60°C until dissolved before use.
3:Homogenize in the FastPrep® Instrument for 40 seconds at a speed setting of 6.0.
4:Centrifuge at 12,000 x g for 5minutes to pellet debris.
5:Transfer supernatant to a clean 2.0 ml centrifuge tube. Add 250μl PPS Solution and mix by shaking the tube by hand 10 times.Incubate at 4°C for 5 minutes.
6:Centrifuge tubes at 10,000 x g for 3 minute at room temperature.Avoiding pellet, transfer up to, but no more than, 900 μl of supernatant to a clean 2 ml centrifuge tube.
7:Add 300 μl of IRS Solution(1/3 volume) and vortex briefly. Incubate at 4°C for 5 minutes.
8:Centrifuge tubes at 10,000 x g for 1 minute at room temperature. Avoiding pellet, transfer the supernatant into a clean 5 ml centrifuge tube.
9:Add 1.5 volumes of PQ Solution to the cleared supernatant and mix by pipetting.
Example: To 1100 μl lysate add 1650 μl PQ Solution. Reduce the amount of PQ Solution accordingly if less supernatant is recovered. A precipitate may form after the addition of ethanol but this will not affect the procedure.
Note: Ensure ethanol has been added to PQ Solution.
Note: It is important to pipet PQ Solution directly onto the cleared supernatant and to mix immediately.
10:Load approximately 700 μl mixture onto Spin Filter(sitting in collection tube) and centrifuge at 10,000 x g for 1 minute at room temperature. Discard flow through. Load another 700 μl and repeat untill all remaining mixture is loaded on Spin Filter.
Note: A total of 4-5 loads for each sample processed may be required.
11:Add 600 μl of Buffer WB to Spin Filter and centrifuge at 10,000 x g for 30 seconds at room temperature. Discard flow through. Repeat Step 11 with another 600 μl Buffer WB.
Note: Ensure ethanol is added to Buffer WB.
12:CentrifugeSpinFilterat13,000xgfor2minuteatroomtemperature to dry the Spin Filter.
13:Carefully place Spin Filter in clean 1.5 ml centrifuge tube. Avoid splashing any Buffer WB onto Spin Filter. Add 100 μl of Elution Buffer (Optional: pre-warm the water to 70–90C will increase the DNA yield) to the center of the column membrane. Incubate at room temperature for 3-5 min, and centrifuge at 13,000 x g for 1 min to elute the DNA.
Note: Use smaller volume(minimum 30μl) of Elution Buffer will obtain higher concentration.
Optional: Put eluate back to the Spin Column to repeat elution once. This increases concentration of DNA about 10-15%.
1:Add up to 500 mg of soil sample to a Bead Tube.
2:Add 980 μl Sodium Phosphate Buffer to sample in Bead Tube.Gentle votex to mix. Add 120 μl MT Buffer.
Note: Check MT Buffer. If MT Buffer is precipitated, heat solution to 60°C until dissolved before use.
3:Homogenize in the FastPrep® Instrument for 40 seconds at a speed setting of 6.0.
4:Centrifuge at 12,000 x g for 5minutes to pellet debris.
5:Transfer supernatant to a clean 2.0 ml centrifuge tube. Add 250μl PPS Solution and mix by shaking the tube by hand 10 times.Incubate at 4°C for 5 minutes.
6:Centrifuge tubes at 10,000 x g for 3 minute at room temperature.Avoiding pellet, transfer up to, but no more than, 900 μl of supernatant to a clean 2 ml centrifuge tube.
7:Add 300 μl of IRS Solution(1/3 volume) and vortex briefly. Incubate at 4°C for 5 minutes.
8:Centrifuge tubes at 10,000 x g for 1 minute at room temperature. Avoiding pellet, transfer the supernatant into a clean 5 ml centrifuge tube.
9:Add 1.5 volumes of PQ Solution to the cleared supernatant and mix by pipetting.
Example: To 1100 μl lysate add 1650 μl PQ Solution. Reduce the amount of PQ Solution accordingly if less supernatant is recovered. A precipitate may form after the addition of ethanol but this will not affect the procedure.
Note: Ensure ethanol has been added to PQ Solution.
Note: It is important to pipet PQ Solution directly onto the cleared supernatant and to mix immediately.
10:Load approximately 700 μl mixture onto Spin Filter(sitting in collection tube) and centrifuge at 10,000 x g for 1 minute at room temperature. Discard flow through. Load another 700 μl and repeat untill all remaining mixture is loaded on Spin Filter.
Note: A total of 4-5 loads for each sample processed may be required.
11:Add 600 μl of Buffer WB to Spin Filter and centrifuge at 10,000 x g for 30 seconds at room temperature. Discard flow through. Repeat Step 11 with another 600 μl Buffer WB.
Note: Ensure ethanol is added to Buffer WB.
12:CentrifugeSpinFilterat13,000xgfor2minuteatroomtemperature to dry the Spin Filter.
13:Carefully place Spin Filter in clean 1.5 ml centrifuge tube. Avoid splashing any Buffer WB onto Spin Filter. Add 100 μl of Elution Buffer (Optional: pre-warm the water to 70–90C will increase the DNA yield) to the center of the column membrane. Incubate at room temperature for 3-5 min, and centrifuge at 13,000 x g for 1 min to elute the DNA.
Note: Use smaller volume(minimum 30μl) of Elution Buffer will obtain higher concentration.
Optional: Put eluate back to the Spin Column to repeat elution once. This increases concentration of DNA about 10-15%.
组分:
组分名称 | 50T | 储存 |
Bead Tube | 50EA | 室温 |
Sodium Phosphate Buffer | 50mL | 室温 |
MT Buffer | 6mL | 室温 |
PPS Solution | 13mL | 室温 |
IRS Solution | 15mL | 室温 |
PQ Solution | 35mL(首次使用前添加两倍体积的乙醇) | 室温 |
Buffer WB | 13mL(首次使用前添加四倍体积的乙醇) | 室温 |
Elution Buffer | 15mL | 室温 |
DNA Bind Columns | 50EA | 室温 |
保存条件:
室温运输,按说明书保存 。
自备试剂:
无水乙醇(E20003)
UN码:
HazardClass:
危害声明:
安全说明:
搜索质检报告(COA)
参考文献 & 客户发表文献
本计算器可帮助您计算出特定溶液中溶质的质量、溶液浓度和体积之间的关系,公式为:
质量 (g) = 浓度 (mol/L) x 体积 (L) x 分子量 (g/mol)
摩尔浓度计算公式
用本工具协助配置特定浓度的溶液,使用的计算公式为:
开始浓度 x 开始体积 = 最终浓度 x 最终体积
稀释公式
稀释公式一般简略地表示为:C1V1 = C2V2 ( 输入 输出 )