Real-Time RT-PCR, which stands for Real-Time Reverse Transcription Polymerase Chain Reaction, combines two well-known techniques, reverse transcription and polymerase chain reaction (PCR), to detect and quantify RNA expression levels in cells. This technique is highly sensitive and can detect even small changes in gene expression, making it a valuable tool for researchers studying gene regulation and disease mechanisms.
One of the most significant advantages of Real-Time RT-PCR is its ability to provide quantification of RNA expression levels in real-time. By using fluorescent dyes which emit light when binding to the amplified DNA, researchers can measure the amount of amplified product at each cycle of PCR, thus providing real-time information on gene expression levels.
RT-PCR is a reaction in which RNA is reverse transcribed into complementary DNA (cDNA) using reverse transcriptase enzyme. The resulting cDNA can then be amplified using PCR to generate multiple copies of the same DNA sequence. Real-Time RT-PCR is one of the most effective and widely used methods of analyzing gene expression.
Two types of Real-Time RT-PCR procedures are used, one-step and two-step. In the one-step RT-PCR procedure, cDNA synthesis and PCR amplification occur in a single tube or well, while in the two-step protocol, a separate cDNA synthesis reaction is performed before the PCR amplification.
When it comes to selecting the right Real-Time RT-PCR kit, there are several factors to consider. The type of RNA being analyzed, the amount of RNA input, and the desired level of sensitivity are all important considerations. Additionally, if you anticipate switching between analyzing mRNA and DNA, choosing a kit that is compatible with both is essential.
A SYBR Green RT-PCR kit for the RotorGene 6000 5 Plex HRM RotorGene is an excellent choice for researchers interested in detecting mRNA in their cells. These kits are highly sensitive and efficient, providing accurate real-time results. However, it is essential to consider the compatibility of the kit with the type of RNA being analyzed and the desired sensitivity level.
If you plan to switch between analyzing mRNA and DNA, it's important to choose a kit that is compatible with both types of nucleic acid. Certain kits, such as the one-step SYBR Green RT-PCR kit, can be used to analyze both DNA and mRNA samples.
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In conclusion, Real-Time RT-PCR is a powerful tool for researchers interested in studying gene expression and disease mechanisms. When selecting the right kit, it's important to consider the type of RNA being analyzed, the amount of input RNA, and desired sensitivity level. Additionally, choosing a kit that is compatible with both mRNA and DNA analysis can save time and resources. TIANGEN BIOTECH (BEIJING) CO., LTD. is a reputable company that offers various services to help customers get nucleic acid from various samples and complete downstream detections, making it an excellent choice for researchers seeking reliable and effective solutions to their research questions.