为什么我的NanoDrop分光光度计或A260/A280值比Qubit荧光计显示的浓度高?
<p>分光光度计不能区分完整核酸、小片段、降解样本,以及单核苷酸。试剂盒中的染料仅检测长度超过20mer左右的完整核酸链。试剂盒仅识别完整DNA或RNA,而忽略降解样本。</p>
Qubit™ 4 Fluorometer
The Qubit 4 Flurometer is the latest version of the popular Qubit fluorometer designed to accurately measure DNA, RNA, and protein quantity, and now also RNA integrity and quality, using the highly sensitive Qubit assays.
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The Qubit 4 Flurometer is the latest version of the popular Qubit fluorometer designed to accurately measure DNA, RNA, and protein quantity, and now also RNA integrity and quality, using the highly sensitive Qubit assays.
The Qubit 4 Fluorometer was re-engineered to run the Qubit RNA IQ (integrity & quality) assay. The Qubit 4 Fluorometer and RNA IQ Assay Kit work together to accurately distinguish intact from degraded RNA in just two easy steps.
For all Qubit assays, the concentration or quality of the target molecule in the sample is reported by a fluorescent dye that emits a signal only when bound to the target, which minimizes the effects of contaminants on the result. The easy-to-use touch-screen menus make it easy to select and run the assays you need, with results displayed in just a few seconds.
Key features of the Qubit 4 Fluorometer include:
The Qubit 4 Flurometer can also be used to directly measure the fluorescence of samples using Fluorometer mode. Additionally, Ion Sphere Particle quality can be evaluated on the Qubit 4 Fluorometer using the Ion Sphere Quality Control Kit prior to performing a sequencing run on the Ion PGM sequencer.
Enables more accurate results than with UV absorbance at low concentrations
The Qubit 4 Fluorometer is far more sensitive than UV absorbance, which measures anything absorbing at 260 nm, including DNA, RNA, protein, free nucleotides, or excess salts. Moreover, UV spectrophotometry is often not sensitive enough to accurately measure low concentrations of DNA and RNA. With the Qubit fluorometer, your research is enhanced by more accurate measurements because the dyes in the Qubit assay kits fluoresce only when bound to the selected molecule (DNA, RNA or protein) in your sample. This allows you to avoid repeating work due to inaccurate measurements.
Fast, simple, and accurate measurement of RNA quality
The Qubit RNA IQ assay provides a fast, simple method to check whether an RNA sample has degraded. The assay utilizes two unique dyes: one that binds to large, intact RNA, and another that selectively binds to small, degraded RNA. Together they enable you to quickly assess the quality and integrity of an RNA sample. To use, simply add your samples to the RNA IQ working solution, then measure on the Qubit 4 Fluorometer.
Save samples and bench space
The thoughtfully designed Qubit 4 Fluorometer requires samples of only 1–20 μL. It is intended for use at room temperature and occupies only a small amount of bench space. The Qubit 4 Fluorometer features advanced optics and data analysis algorithms, and comes equipped with a USB thumb drive and cable for data transfer to Excel™ software and for software downloads, as well as a universal power supply, four plug adapters, and electromagnetic CE certification.
Single sample measurements are a breeze
This user-friendly fluorometer is combined with quick and simple assay procedures. Calculations and settings are automatically performed by the instrument. The Qubit assays for use with the Qubit 4 Fluorometer are all performed using the same general protocol, which uses a simple mix-and-read format with incubation times of only two minutes for the DNA and RNA assays. 500μL thin-walled PCR tubes (Cat. No. Q32856) are required.
System specifications include:
采购问答
<p>分光光度计不能区分完整核酸、小片段、降解样本,以及单核苷酸。试剂盒中的染料仅检测长度超过20mer左右的完整核酸链。试剂盒仅识别完整DNA或RNA,而忽略降解样本。</p>
<p>请从我们的网站下载并安装最新版本的Qubit固件(https://www.thermofisher.com/us/en/home/industrial/spectroscopy-elemental-isotope-analysis/molecular-spectroscopy/fluorometers/qubit/qubit-technical-resources.html)<br/><br/> 对Qubit 1.0, 2.0 和 3.0荧光计,去这个链接下载:https://www.thermofisher.com/us/en/home/industrial/spectroscopy-elemental-isotope-analysis/molecular-spectroscopy/fluorometers/qubit/qubit-technical-resources/previous-qubit-models.html </p>
<p>对于Qubit 1.0仪器,如果它和电脑相联,那么将它和电脑的连接解除,拔掉电源,等待至少10秒,然后重新插上电源。<br/><br/> 对于Qubit 2.0,3.0,4.0 仪器,如果可能,关闭电源或拔掉电源,等待至少10秒,然后重新插上电源。在固件升级过程中,请不要将仪器电源拔掉或将USB闪存拔掉。</p>
<p>如果可能,你可以试试使用较大体积的样本(最多20µL)。此外,如果你使用的是Qubit DNA BR检测试剂盒或Qubit RNA BR检测试剂盒,可以试试Qubit DNA HS检测试剂盒或Qubit RNA HS检测试剂盒,因为它们的灵敏度更高。</p>
<p>稀释你的样本,或者使用较小体积的样本,重新检测。此外,如果你使用的是Qubit DNA HS检测试剂盒或Qubit RNA HS检测试剂盒,可以试试Qubit DNA BR检测试剂盒或Qubit RNA BR检测试剂盒,因为它们的灵敏度更低。</p>
<p>如果出现这种情况,在“查看标准品”或"查看校准"下查看标准品和样本的原始荧光值。确认样本的荧光值落在标准品的荧光值之间(或稍稍高于最高的标准品)。如果不是,那么样本已经超过了检测的精确范围。Qubit荧光计的屏幕会显示“超出检测范围”。</p>
<p>这条信息出现可能有几个原因。请在您的仪器上检查如下事项:<br/><br/> 1.在 “查看标准品”或"查看校准"下查看标准品的原始荧光值。Standard 2的荧光值是否至少是Standard 1荧光值的10到50倍。对于蛋白检测,Standard 3的荧光值是否是Standard 2荧光值的大约2倍。<br/> 2.检查你的标准品放置顺序是否正确。<br/> 3.检查你是在合适的检测管中使用了10µL的标准品和190µL的Qubit染料工作溶液。<br/> 4.对于RNA检测,可以试试新开一管Standard 2,以防已经打开的RNA标准品已经降解。<br/> 5.检查Qubit检测试剂盒以及标准品的收货日期。试剂盒从收到的日期起可以稳定使用至少6个月。如果该试剂盒已经超过一年并且Qubit荧光计不能读取标准品,那么应当换用一个新的试剂盒。<br/> 6.检查仪器是否正常工作。打开盖子。运行任一Assay的新的校准。你可以看到一个红色灯(对于RNA Assay)或一个蓝色灯(对于所有其它Assay)闪烁5次。</p>
<p>可以,使用手册中有关于此应用的说明。</p>
<p>是的。我们观察到20–30%的差别。对于不同形态的质粒DNA,我们建议使用和样本中的质粒DNA形态非常近似的标准品。欲获知创建自定义优化检测法的信息,请访问我们的网站(https://www.thermofisher.com/search/results?query=MyQubit)。</p>
Sende Lab
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<p>是的。对于典型的高浓度的小抽DNA(超过50 ng/μL),可以使用Qubit DNA BR检测法。对于微量质粒或少量质粒,可以使用Qubit DNA HS检测法。但是,关于构象问题请参见下一问题。</p>