We learned about the huge variety of both coding and noncoding type...

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We learned about the huge variety of both coding and noncoding types of RNA that cells produce and a great deal about their natural instability and susceptibility to ribonuclease (RNase) assault. We know that RNA can serve as a template for the synthesis of cDNA and, if desired, that the cDNA can be amplified by PCR. Please explain which of the following milestones you believe has had the biggest impact on the study of gene expression, and why. The choices are:
(a) ability to isolate intact RNA from cells and tissues,
(b) the ability to synthesize cDNA,
(c) the development of PCR, or
(d) if you don't like these milestones, then feel free to proffer something else (be sure to thoroughly explain your thinking).

Second, to respond to the following three postings, in which you agree with them or disagree with them, and explain why. Your responses should be well thought out, consist of at least a paragraph or two,, be courteous, and be respectful.

First post
As per my belief in the study of gene expression the development of q PCR and cDNA synthesis from mRNA are the biggest effective things which highly impact on current study going on gene expression. The reason behind this is as follows-Gene expression analysis helps in the study to unpack how genes are regulated, and how gene expression changes in different conditions and at different stages of lifecycles, for example during cell division, in disease states in humans and animals, or during disease and pathogen-defense responses in plants. This includes when and which genes are suppressed or blocked, expressed and overexpressed.

Applications for gene expression analysis contains basic biological research in developmental biology, Understanding cell division and cell pathways, Quantitation of RNA and gene products siRNA/RNAi experiments, Quality control and assay validation, Microarray validation, Phenotypic analysis of cells and tissues, Gene expression changes in disease, PCR in gene expression profiling.etc.

Gene expression is generally measured in terms of expression of messenger RNA (mRNA), which is transcribed from DNA and then used as a template to code for proteins. However, proteins are not the only gene products, and assays are being developed to profile microRNAs (miRNAs), which have regulatory roles, and other non-coding forms of RNA, including ribosomal RNA (rRNA) and transfer RNA (tRNA).

In gene expression profiling the mRNA is isolated and converted to cDNA (complementary DNA) using reverse transcription. The cDNA fragments are then amplified using qPCR and the quantity and proportions of the original RNA calculated based on the fluorescent signals received.

Second post
I believe that the ability to synthesize cDNA has had the biggest impact on the study of gene expression. There are a ton of advantages when it comes to synthesizing cDNA. The mRNA being converted into cDNA goes from being one-stranded to a more stable one-strand or even two stranded DNA. More storage of amino acid sequences for gene expression is the benefit here. Ligation can be used to create more cDNA as needed. cDNA cannot be made from mRNA that the cell is not producing, which means its noted in active gene expression. One of the last key advantages of cDNA is that it can be linearly amplified to produce more RNA relative to the amount in the cell. The only real important weakness of using cDNA is that it is not done in the cell, but in-vitro. cDNA is also synthesized directly from RNA which allows for gene expression. cDNA varies depending on the tissue it comes from, which means it produces specific amino acids for different functions. This is different from gDNA. There are also many different kinds of enzymes such as RNase H and DNA polymerase 1 which synthesis cDNA. Since there are a bunch of enzymes that can synthesis cDNA, this allows for more and easier transcription from mRNA to cDNA making it more important for gene expression.

Third post
When it comes to gene expression all of these three choices are essential in gene expression “central dogma” process because they complete each others. From the research I have done and what I have been learning so far I think that RNA has the biggest impact on gene expression and that is because of how useful and important RNA is when it comes to gene expression. To start off, gene expression is a process where information from a gene is used in the synthesis of a functional gene product. These products are often proteins, but in non-protein coding genes such as transfer RNA or small snRNA genes. Therefore, RNA plays many roles in and during gene expression which helps us to study it better. RNA plays an essential role in each step of the gene expression process. It starts with the DNA molecule which contains a gene that is transcribed into RNA.Then, we go in how mRNA exits the nucleus into the cytoplasm. Lastly, the RNA is translated into protein by matching the right amino acid with its cognate RNA codon. However, the different RNA molecules plays a role in these processes. The RNA molecule is transcribed from

DNA by an enzyme which is known as RNA polymerase. In regulation of RNA to gene expression what happens is that a set of transcription proteins gets tighter and bind to a specific DNA sequence that is near the gene and promote the transcription into an RNA.In a number of clinically important viruses RNA, rather than DNA, carries the viral genetic information. RNA also plays an important role in regulating cellular processes–from cell division, differentiation and growth to cell aging and death. In conclusion, without RNA it would be difficult to study gene expression but not only RNA that is needed in a process like this. PCR is also important because is used in the sciences filed for things like disease diagnosis and forensics science.

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My Answer:
I believe that the development of PCR has been the biggest impact on the study of gene expression, for the simple reason that it is the single most influential technique of molecular biology that has impacted all kinds of research subsequent to its innovation in 1986-87 by Kary B. Mullis and Fred.A. Faloona. Prior to its invention, DNA dependent DNA polymerase was already known by Kornberg et al, the central dogma of molecular biology has already been reversed by the discovery of Reverse transcription by Temin et al, and Sanger’s di-deoxy method was developed. Scientists were able to amplify DNA by cloning through plasmid vectors; however, they didn’t have any way to amplify any sequence of DNA in vitro, directly from a sample without any hectic procedure. Through PCR they could amplify even a single piece of DNA to about a billion times in just a few hours. Expression of genes could still be analyzed through other techniques such as protein profiling with Western blotting prior to the development of PCR and so as the analysis of RNA via Northern and radioactive probes. However, PCR paved the way for time-saving, amplification of nucleic acid that revolutionized the whole molecular biology field. It's true that RNA dependent DNA polymerase or reverse transcriptase and the subsequent development of cDNA synthesis was a landmark, however, it was incomplete in itself to mark an impact on gene expression analysis, until combined with techniques such as PCR to eventually develop into Real-Time PCR and other technology....

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