PCR Clean™的科研实验数据

Testing PCR Clean™ Efficiency

PCR Clean的科研实验数据

INTRODUCTION

PCR Clean™ is a ready-to-use solution for the removal of nucleic acids from any surface at PCR workstations and/or lab devices and equipment. The solution contains a surfactant and a non-alkaline and non-carcinogenic agent. In this study we examined the efficiency and effectiveness of PCR Clean™ against different nucleic acid contaminations on various surfaces, in comparison with several other cleaning agents.

介绍

PCR Clean™是一款即用型工作液，用于去除PCR工作台、实验室机电产品和设备表面上留存的核酸。该工作液包含一种表面活性剂和一种非碱性非致癌性试剂。在这项研究中，我们针对不同的表面上留存的不同类型的核酸，检查了PCR Clean™的使用效果和效率，并和其他的几种去除剂进行了比较。

PROCEDURE

1. Removal of amplicon DNA from various surfaces

The efficiency of PCR Clean™ was tested for the removal of bacterial amplicon DNA from E. coli 0104 and E. coli 0157 and compared to several other cleaning materials. Bacterial amplicon DNA was applied by pipetting 2 µl (0.05 ng) E. coli 0104 DNA on a plastic foil, glass surface, and lab work surface area (Trespa ® ), or 2 µl (0.2 ng) E. coli 0157 DNA on Plexiglas ® and aluminum-surfaces. After DNA was completely air-dried, it was removed by using a paper towel moisturized with PCR Clean™, a diluted dishwashing detergent, 70 % Ethanol, Isopropanol, water, or a dry paper towel, by wiping off the spot where DNA was pipetted with one stroke. Samples were then collected by using a moisturized swab and wiping off (swabbing) the same area. As positive control, either 0.05 ng E. coli 0104 DNA (for plastic foil, glass or Trespa ® assays), or 0.2 ng E. coli 0157 DNA (for Plexiglas ® and aluminum assays), were directly pipetted into 250 µl PCR grade water and extracted in the same manner as the other samples.

实验过程

1、去除不同表面上的DNA扩增片段

我们用大肠杆菌O104和O157的DNA扩增片段，检查了PCR Clean™对DNA污染的去除效率，并和其他几种去除试剂进行了比较。我们在塑料薄膜、玻璃表面和千思板（常用于实验工作区台面，Trespa®品牌）滴加了2μl（0.2 ng）大肠杆菌O104 DNA，在Plexiglas®（树脂玻璃）和铝表面滴加了2μl（0.2ng）大肠杆菌O157 DNA。待DNA完全风干后，用蘸有PCR Clean™或一种稀释的洗洁精、70%乙醇、异丙醇、水的纸巾擦拭去除，或者用干纸巾擦拭去除。随后用一个干净潮湿的拭子在该区域表面涂拭取样。另取0.05 ng大肠杆菌O104 DNA或0.2 ng大肠杆菌O104 DNA作为阳性对照，加入250l PCR级别水溶解，其抽提方式和其他样品一样。

2. Removal of genomic DNA and plasmid DNA from aluminum surfaces

The efficiency of PCR Clean™ was also tested for genomic and plasmid DNA removal from aluminum surfaces by pipetting 2 µl (2 × 10 6 genome copies) of E.coli genomic DNA, or 2 µl (2 × 10 6 genome copies)

2、去除铝表面的基因组DNA和质粒DNA

在铝表面滴加2μl（2X10⁶拷贝数）大肠杆菌基因组DNA或质粒DNA测试。PCR Clean™与蘸水的湿纸巾或干纸巾对照。

3. Removal of RNA from a Plexiglas ® surface

5 µg (11 µl) were pipetted on each of 6 different spots of a Plexiglas ® surface.

3、去除Plexiglas®（树脂玻璃）表面上的RNA

在Plexiglas®表面6个点分别滴加5g（11μl）RNA进行测试。取样后的RNA作逆转录和qPCR。

4. DNA extraction and qPCR amplification

DNA extraction from the swabs was carried out using our SwabUp™ Lab Monitoring kit. DNA was extracted from all

collected samples and positive controls in the same manner. All DNA extracts were subsequently subjected to qPCR.

4、DNA抽提和qPCR扩增

拭子上的DNA采用SwabUp™ Lab Monitoring试剂盒进行抽提。所有收集的样本DNA和阳性对照抽提的方式相同。随后进行qPCR。

RESULTS

1. Removal of amplicon DNA from various surfaces

Results showed a bigger depletion of amplicon DNA after removal with PCR Clean™ from the various surfaces tested, in comparison to other cleaning agents and in reference to the positive control (Table 1). For all surfaces tested, PCR Clean™ showed the best effectiveness for the removal of amplicon DNA, which could be observed in Graph 1, where Ct -values for PCR Clean™ were higher in comparison to other cleaning agents and in reference to the positive control, regardless of the surface material (Graph 1, A-E), indicating a higher decrease in DNA amount.

检验结果

1、去除不同表面的DNA扩增片段

结果显示，和其他几种清除剂相比，PCR Clean™的对扩增的DNA片段去除的更多（见表1），效果**（见图1），Ct值更高。

表1：与其他清除剂相比，使用PCR Clean™后，qPCR测定的细菌DNA扩增片段的Ct值

图1：采用不同的清除剂，对 A.树脂玻璃®、B.铝、C.千思板®、D.玻璃和E.塑料薄膜上的大肠杆菌DNA扩增片段去除后，qPCR的扩增曲线

2. Removal of genomic DNA and plasmid DNA from aluminum surfaces

Results showed a bigger depletion of genomic DNA and plasmid DNA after removal with PCR Clean™ from an aluminum surface

2、去除铝表面的基因组DNA和质粒DNA

结果显示，与水和干纸巾相比，含PCR Clean™的湿巾对基因组DNA和质粒DNA有更多的去除（见表2）。Ct值（图2,A，B）更高，提示DNA量有大幅下降。

表2：与水和干纸巾相比，使用PCR Clean™后，qPCR测定的铝表面基因组DNA或质粒DNA的Ct值

图2：采用PCR Clean™、水或干纸巾去除铝表面的大肠杆菌基因组DNA或大肠杆菌质粒DNA后的qPCR扩增曲线

3. Removal of RNA from a Plexiglas ® surface

Results showed a bigger depletion of RNA after removal with PCR Clean™ from a Plexiglas ® surface

3、去除Plexiglas®（树脂玻璃）表面的RNA

和其他几种清除剂相比，PCR Clean™对Plexiglas®表面的RNA去除最多（见表3），Ct值也最高（见图3）

表3：和其他几种清除剂相比，PCR Clean™去除树脂玻璃表面的RNA后，RNA逆转录合成cDNA, 再qPCR测定Ct值

图3：使用不同的清除剂，从树脂玻璃®中去除RNA后，RNA再逆转录并进行qPCR扩增

CONCLUSIONS

The results of our study showed that PCR Clean™ is highly effective against amplicon, plasmid, and genomic DNA, and RNA contaminations from all surfaces tested, even within seconds after use. Depletion results also demonstrate superiority in effectiveness and efficiency of PCR Clean™ compared to any of the other common cleaning agents usually available at molecular biology laboratories, e.g. ethanol, isopropanol or dishwashing detergent. Therefore, the regular use of PCR Clean™, both before and after PCR analysis is fast, easy and ideal to maintain a clean work area, which can be critical in molecular biology laboratories and PCR workstations, and thereby saves time and expenses.

结论

我们研究的结果显示，PCR Clean™对来自所有检测表面的DNA扩增片段，质粒、基因组DNA和RNA污染的去除，是极为有效的，而且起效很快。去除结果还显示，相比于其他常见的DNA清除剂，比如分子生物学实验室常用的乙醇、异丙醇或洗洁精，PCR Clean™的去除效果和效率都具有明显的优势。因此，在PCR反应前后，经常使用PCR Clean™，会非常便捷，可理想地保持干净的工作区域，而这对于分子生物学实验室和PCR工作台都是非常重要的，因为它可节省时间和支出。