The application of RNA isolation strategies in widespread use. The method of "grabbing the RNA" depends on the biological source: cells grown in culture versus tissue samples, or plant versus animal tissue, as two examples.
Now, think about cancer in humans. Give some thought to whether it is better to study a previously unknown type of cancer using cell culture or using tissue obtained by biopsy. Be sure to give very specific examples of (a) how your approach could provide new information, and (b) how you would plan to recover the RNA from the biological source. You can, and should, draw on some of the information that we've covered in this lesson and in previous lessons to develop your answer. In the unlikely event that you are unfamiliar with some of the properties of cancer cells, please take a look at the attached summary document, which may help you with the thought process.
Be sure 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 few sentences, be courteous, and be respectful.

First post
I think that using a cell culture would probably be better than using a tissue obtained by biopsy because it would be easier to work with. Cell cultures have a large surface area that is exposed which makes them easier and more efficient to lyse. On the other hand, tissues are not easy to disrupt, or obtain the RNA from. In addition to that, tissues are not easy to obtain, while cell culture can be grown in a lab and are abundant to use in different experiments and studies. With my approach, since there will be more cells to use for different experiments, more options can be available to try. In other words, if one experiment does not work, then there will always be a chance to try a different one until a result is shown. A way to recover RNA would be by using Silica Technology which allows the RNA to bind to silica in a high salt environment. Eventually, small elution volumes will be produces which are favored since the sample will be more concentrated.

Second post
I will choose to study a previously unknown type of cancer using tissue obtained by biopsy. I could look to study the gene expression of biochemically different organisms. This would be especially relevant for my personalized medicine preparations. (a) This approach will provide new information about how the cancerous cell RNA differs from RNA in healthy cells. The difficulty of using biopsy tissue will be worth it for me to use my findings for research purposes. I will be able to look for both subtle and not as subtle biochemical changes that accompany the cancerous cell. I hope to see the very disorganized nature of the cancerous cell gene expression compared to the normal cells in the tissue. (b) I would plan to recover the RNA from the biological source by first choosing a lysis buffer for nuclear RNA. The lysis buffer I choose will be “harsh” guanidinium buffer with RNase inhibitors not added. Nuclease activity can be inhibited without the addition of the inhibitors with this lysis buffer. The sample will be kept at an appropriate temperature, around -80 degrees, through flash-freezing the sample in liquid nitrogen. I would remove protein from the sample next. I would prefer to use an organic solvent to chemically remove them. I would choose to bind RNA reversibly on a silica glass fiber column and then elute it in a small volume. I choose this approach because as noted in the module silica-based procedures are in widespread use. I could draw from the knowledge on a more widely practiced approach. The RNA will need to be stored properly to prevent degradation. I hope to store the purified RNA at -80 degrees as well through the liquid nitrogen approach.

Third post
researching cancer cells is not something that can be determined one way, if I was to study cells I would do both biopsy tissue and cell culture because they each have something to offer that other may not. when using tissue obtained by a biopsy, the cells are exactly how they would be found in the body, they have the specific mutations that originally caused the cancer. the tissue sample, depending on size, has the ability to yield a vast amount of RNA. the more RNA that is able to be isolated means there is more chances to discover something new. taking a biopsy from someone also allows for more personalization of treatment, cancer as we know is not all the same, different mutations and environmental factors cause differences that cell culture would not be able to show. but on the flip side, cell culture allows for different tests to be done. cell cultures are easier to obtain RNA samples because there is typically less of them on the plate. cell cultures are also from specific cell lines that are easy to get and no need for a patient to have a biopsy. cultured cells are also better when specific RNA is of interest, like mitochondrial RNA or cytoplasmic RNA, unlike tissue samples where it is usually total RNA from all cellular compartments. therefore if I was to research a cancer I would use both cell culture and tissue biopsy.

The application of RNA isolation strategies in widespread use. The...