What Are Downstream Applications and RNA?
Downstream applications are references to a direction as it applies to the genetic codes in RNA or DNA strands. It is one of the premier manipulation steps beginning with a biological sample. The process involves creating a DNA microarray sample intended for genotyping and further analytical purposes. The upstream application process is directed toward the upper 5’ end of the DNA or RNA strand. The downstream process is pointed to the lower portion of the DNA or RNA strand which is 3’ long.
These represent a part of the transcription process by which DNA is copied into RNA by polymerase enzymes. Both DNA and RNA are nucleic acids, however, RNA is the intermediator between DNA and Protein. The main distinguishing component is that RNA is a single strand that contains ribose and the nucleobase uracil. DNA also a nucleotide only contains deoxyribose and the nucleobase thymine. Extraction of DNA, RNA, and protein are the hallmark methodology used in genetic laboratories. The extracted components can be isolated from any biological material to be used for genotyping, research, and other downstream application processes.
Geneticists Use Downstream Applications to Isolate Micro-RNA (miRNA)
Various methods of downstream applications are used in genetic laboratory practices to extract and isolate miRNA biomarkers for analyzing specific expressions. Downstream applications are one of the most utilized components in the creation of TMAS (Tissue Microarray Samples, generated by Geneticist Inc. ) commonly used in the scope of molecular biology and research practices. Creating an applicable work-flow is used as a sort of a template that may be used in an SOP as a duplication model in the lab.
About Micro-RNA (miRNA) - miRNAs are a small class of noncoding RNA molecules of which consist of 22 nucleotides that exist in plants, animals and some viruses. They function in RNA silencing and post-transcriptional gene regulation. This has been especially valuable in its role in helping to understand certain cancers such as colorectal cancer. Unfortunately, in 80% of all colorectal cancer cases, it has metastasized before diagnosis, thereby making treatment options ineffective. A study detailed research of miRNA and its role in affecting the effectiveness of gene regulation. The research is centered around understanding this role at the molecular level in order to speed up diagnostic times and assist in improving patient outcomes
Using Downstream Applications to Isolate Circulating Tumor Cells - Guiding Targeted Cancer Interventions and Therapies
CTC’s are rare cancer cells released from tumors into the bloodstream. CTC’s are thought to be one of the most telling byproducts in predicting metastasized tumors in most major forms of cancers such as breast and colorectal. There is a large interest in researching CTC’s as a potential source of cancer biomarkers to assist with the diagnosis and prognosis of various cancers. Research may also facilitate drug development and potential adjuvant therapies, however, there are several challenges as they still seek to determine Pharmakinetics and toxic potential for the miRNA targeted replacement therapies.
Efficacy in the chosen downstream application used to isolate CTC’s comes into strong play in order to help researchers gain access. The isolation methods must be sensitive enough to the rare heterogeneous profile of the CTC population making it a challenging process. Successful clinical applications involve the development of superior analytical tools to assist in affecting potential outcomes. Each downstream application has its own recommendations for:
Cell Purity
Sensitivity
Cell Recover Following Enrichment
Characterization Method for False Positives and Negatives
How Much of a Role Does Downstream Equipment in Choosing the Downstream Application?
The answer to this question is that equipment does play a significant role. If the equipment is fairly complex, the downstream application method is affected. The overall downstream process involves recovering a biochemical product from a series of a mixture of impurities and solvents. The operation will physically change the biological product at every step. The process details can include:
Separation
Isolating levels of concentration
Purification
Distillation
Chromatography
Use of a Biocatalyst
Buffering
Essentially, the downstream application process is an expensive one that requires a biochemical engineer to steer the process from start to finish. SOP’s (Standard Operating Procedures) assist in eliminating costly mistakes in handling. A genetic biorepository is responsible for the safe handling of all specimens and providing the product specified for analytical purposes. All lab procedures including downstream or upstream applications are selected with the overall best practices in excellence and reliability. The scientific result is completely dependent upon the integrity of the isolated nucleic acid.
Conclusion
Downstream applications in genomic laboratory function play a large role in facilitating research. Geneticist Inc. provides human tissue samples for analytical research in various capacities. As one of the leading biorepositories, our use of downstream applications largely dependent upon the type of equipment used, combined with effective methodology and SOP management. Our facility understands the importance of the quality of our product for its intended use in analytical research.
