Why not try a microRNA array to determine which miRNAs are present and highly expressed?

Ryan

Read the following paper
* Tables


Abstract

Eukaryotic cells express small noncoding RNAs to silence target genes or regulate chromatin domains. MicroRNAs (miRNA) are endogenous small RNAs that are thought to each regulate multiple mRNA targets. To gain a deeper understanding of processes regulated by small noncoding RNAs, techniques are required to identify and detect them. This unit describes standard laboratory methods for and comments about high-throughput technologies for the identification and detection of small RNAs. This unit also outlines a cell-based reporter gene assay for assessing the regulatory potential of a microRNA on a candidate mRNA target. Curr. Protoc. Mol. Biol. 81:26.7.1-26.7.15. 2008 by John Wiley & Sons, Inc.

ryan_m said:

Why not try a microRNA array to determine which miRNAs are present and highly expressed? Ryan

Thank you for reading my topic and giving me an advice! However, My boss wouldn't spend much money in setting up a method! He asked me to select a cell line and a target microRNA which has been verified! Finally,I am determined to select A549(a kind of lung cancer cell lines) and hsa-miR-21 (highly expressed) !Then isolating microRNA and performing real-time PCR to check it!
I don't know whether such design is OK!
So could you give me some better advices?
Another question :if I want to use SYBER GREEN to perform QPCR,which kit is suitable to select? The SYBER GREEN I type or II type?

saswati1 said:

Read the following paper * Tables Abstract Eukaryotic cells express small noncoding RNAs to silence target genes or regulate chromatin domains. MicroRNAs (miRNA) are endogenous small RNAs that are thought to each regulate multiple mRNA targets. To gain a deeper understanding of processes regulated by small noncoding RNAs, techniques are required to identify and detect them. This unit describes standard laboratory methods for and comments about high-throughput technologies for the identification and detection of small RNAs. This unit also outlines a cell-based reporter gene assay for assessing the regulatory potential of a microRNA on a candidate mRNA target. Curr. Protoc. Mol. Biol. 81:26.7.1-26.7.15. 2008 by John Wiley & Sons, Inc.

Thank you very much for reading my topic!But this paper is not available! Would you please send this article to my email address? My email address is wangmei8417@163.com!