PrimeDirect Probe RT-qPCR Mix is designed for one-step, real-time RT-PCR via probe detection (5′-nuclease method). This product is supplied as a 2X premix to facilitate the easy preparation of reaction mixtures. It requires only the addition of your primer, probe, and sample, and the RT-qPCR can be performed by simply adding your sample directly to the reaction mixture without intervening nucleic acid purification steps.
PrimeDirect Probe RT-qPCR Mix is designed for one-step, real-time RT-PCR via probe detection (5′-nuclease method). This product is supplied as a 2X premix to facilitate the easy preparation of reaction mixtures. It requires only the addition of your primer, probe, and sample, and the RT-qPCR can be performed by simply adding your sample directly to the reaction mixture without intervening nucleic acid purification steps.
The protocol allows all steps, from extraction of the nucleic acid to reverse transcription and qPCR, to be performed in a single tube, greatly reducing hands-on time and risk of contamination. The single-tube workflow also allows the detection of RNA viruses with complex higher-order structures.
High-efficiency PCR after cDNA synthesis detects DNA amplification in real time by measuring increased fluorescence from the probe. Because this product is highly resistant to various PCR inhibitors such as heparin (blood) and humic acid (soil), it can be used to directly detect pathogens, including viruses and bacteria, in various biological specimens and to directly analyze gene expression in cells.
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请问PCR仪设置加样量有什么作用,我们实验室有的pcr仪可以设置有的不能设置。如果我用了25微升体系,pcr仪设置时设置了50微升体系会怎么样呢?帮忙回答一下。谢谢~
2、放 PCR 离心管,先把顶盖向上扳,再往前推,露出放样孔,PCR 管
放入前应加好反应体系各组分,再放入 PCR 离心管。
3、反向重复第二步骤将顶盖板向后拉,盖好顶盖。
4、按 F2“Create”新建一个扩增的 PCR 程序。
5、按控制台面右方上下左右方向键,可随意移动编辑位置,输入需要
的温度、时间、循环数等。
6、据 PCR 离心管中加入的反应体积总量,在“Reaction volume”后输入
相应数值,有 Std“1~100ul”和 9600“1~50ul”两档可供选择。
7、按 F1“Start”启动
8、按“INFO”对应键查看 PCR 结束键。
9、扩增完成后按台面左方“Stop”键退出。
10、按“Hist”,可查看上次扩增的历史记录。11、完全退出后,按台面左下方电源开关,拔出插销,切断电源。