I want to know about the difference between qPCR and RTPCR. What is major difference between these two PCRs? For which experiments, I have to use these PCRs?
RT-PCR is term used for "Reverse Transcription" PCR , where the starting genetic material in the PCR reaction is RNA ,where RNA is first transcribed in reverse into its DNA complement by reverse transcriptase enzyme. The new complementary DNA containing the reversed transcription will then be amplified using the traditional PCR or real-time PCR. and here if you proceed with traditional PCR you will read your results as "presence or absence" of the amplified gene product using agarose gel electrophoresis (No quantification).
While qPCR (Quantative PCR)or sometimes called digital PCR you have a qPCR machine and a fluorescent dye in your PCR reaction (Sybr green or Taqman Probe) that bind to your amplified gene product and give you a Ct value , which later on gives you an idea about the quantity or your RNA or DNA in your reaction depending on a standards used , in this case you are not in need to do gel electrophoresis to see your amplified pcr product.
so qPCR could be also RT-qPCR if you work on RNA or only qPCR if DNA is your starting genetic material.
qPCR is a quantitative PCR using a scale or standard curve. RTPCR is real time PCR and does not imply a quantitative analyses.
RT-PCR (Reverse Transcription-PCR): usually you start from RNA, reverse transcribing to cDNA, then do the PCR reaction (usually to measure gene expression)
qPCR (quantative-PCR): the amount of DNA after each cycle of replication is measured, usually using fluorescent dyes (used to quantitatively measure the amplification of DNA)
This two terms usually make confusion to beginers. In old published paper, they write RT-PCR but it is not reverse transcriptase PCR. They mentioned it as Real Time pcr. However, later, becaise of this confusion, all scientists agreed to write real time PCR as qPCR and RT-qPCR (for reverse). Then, nowadays RT-PCR is simply meant to reverse transcriptase reaction only for normal PCR. Therefore, if you are reading old papers, be careful to know how writer termed his experiment.
RT-PCR is term used for "Reverse Transcription" PCR , where the starting genetic material in the PCR reaction is RNA ,where RNA is first transcribed in reverse into its DNA complement by reverse transcriptase enzyme. The new complementary DNA containing the reversed transcription will then be amplified using the traditional PCR or real-time PCR. and here if you proceed with traditional PCR you will read your results as "presence or absence" of the amplified gene product using agarose gel electrophoresis (No quantification).
While qPCR (Quantative PCR)or sometimes called digital PCR you have a qPCR machine and a fluorescent dye in your PCR reaction (Sybr green or Taqman Probe) that bind to your amplified gene product and give you a Ct value , which later on gives you an idea about the quantity or your RNA or DNA in your reaction depending on a standards used , in this case you are not in need to do gel electrophoresis to see your amplified pcr product.
so qPCR could be also RT-qPCR if you work on RNA or only qPCR if DNA is your starting genetic material.
1. Both QPCR and RT-PCR are used in biotechnology and used for the production of multiple copies of DNA.
2.RT-PCR is used to amplify the reversed transcription of the DNA code where as the QPCR measures the amplification.
3.QPCR is quantitative in nature while RT-PCR is not
Dear Dr. Aqleem
i agree with all respected answers from all that mention above and
i can summery the different between them in these points:-
1.QPCR and RT-PCR are both terms used in biotechnology and utilized for the production of multiple copies of DNA. 2.RT-PCR is used to amplify the reversed transcription of the DNA code; QPCR measures the amplification. 3.RT-PCR is for amplification, while qPCR is for quantification. 4.QPCR is quantitative in nature, while RT-PCR is not.
i suggest you read this
Basic of quantitative real time PCR
Presentation Basic of quantitative real time PCR
good luck to you
RT-PCR can be used without qPCR, for example to enable molecular cloning, sequencing or simple detection of RNA. Conversely, qPCR can be used without RT-PCR, for example to quantify the copy number of a specific piece of DNA.
Mohamed Rasheed Gadalla sir I have bit confusion in one thing, I have to perform on an experiment where I need to quantify MT-DNA from plasma samples of humans and mice disease model, which one should I use and why? for mtDNA analysis, there is no need for performing RT PCR? plz reply.
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