Another epigenetic regulation mechanism is microRNAs, a class of tiny RNA (18-22

Question

Another epigenetic regulation mechanism is microRNAs, a class of tiny RNA (18-22 nts) that regulates various physiology functions including cancer development. Currently microRNAs are being investigated as a new generation of biomarker for cancer early detection. Accurate microRNA quantification is highly demanded for both research and clinic diagnostics. Traditional methods include RT-PCR, microarray, insitu hybridization, and northern blotting (no longer used), and RT-PCR is the gold standard.

Please read the following paper about a microRNA detection method.

Paper 1 to be read:

Areta et al. Rapid microRNA detection using power-free microfluidic chip: coaxial stacking effect enhances the sandwich hybridization. Analyst, 137, 3234 (2012) http://pubs.rsc.org/en/Content/ArticleLanding/2012/AN/c2an16154k#!divAbstract

Two addition references by the same group are provided, if you are interested:

Areta et al. Rapid and Sensitive MicroRNA Detection with Laminar Flow-Assisted Dendritic Amplification on Power-Free Microfluidic Chip. PLoS One. 2012; 7(11): e48329. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3492330/

Areta et al. Rapid Sub-attomole MicroRNA Detection on a Portable Microfluidic Chip. Analytical Sciences, 30(1), 129 (2014). http://www.jsac.or.jp/analsci/abst.php/30/1/129/

So far, we cannot give comment whether this method can be developed into a clinically usable technique. However, we notice that this microRNA sensor utilizes an important molecular mechanism, called coaxial stacking, which is a fundamental interaction involved in DNA/RNA, in particular RNA tertiary structure assembling. Therefore students can learn from this paper about how to design nucleic acids molecular system with enhanced hybridization strength. It is also a basic interaction utilized to fold DNA/RNA nanostructures (DNA/RNA nanotechnology).

1)What is in nucleic acids structure________

2)The three fragments playing in the hybridization are 1. miRNA (, _______bases), 2. Fluorescent labelled probe DNA (, ______bases ), and probe immobilized on glass (, ______bases ).

3)Two detection protocols, Protocol 1 and protocol 2, have been tested. Please fill in the specific fragment names above and their hybrid names in the following procedures:

In Protocol 1, _____ is first hybridized with______, then the ___________hybrid is hybridized with_______

In Protocol 1, _____ is first hybridized with______, then ______is added to hybridize with_______

4)Protocol 1 was found to be 10 fold more sensitive (lower low detection limit) then Protocol 2. Please explain why_____________________________

Explanation / Answer

1)

A long polymeric strands of nucleotides is called nucleic acids, it may be either DNA (double stranded) or RNA (ribonucleic acid). Thus, both RNA and DNA made up of four different types of nucleotides called A, T, G and C. (note: U found in RNA in the place of T.

2)

The three fragments playing in the hybridization are 1. miRNA (18 to 24 bases), 2. Fluorescent labelled probe DNA (11 bases), and probe immobilized on glass (, 29 bases).     

3)

In Protocol 1, miRNA is first hybridized with biotinylated probe DNA then the miRNA and probe DNA hybrid is hybridized with F-SA.

In Protocol 2, miRNA is first hybridized with biotinylated probe DNA then the miRNA and probe DNA hybrid is hybridized with Bio-anti-SA

Related Questions

Navigate

• Browse (All)
• Browse A
• Subjects

Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.