Principles
In RT-PCR, the RNA template is first converted into a complementary DNA using a reverse transcriptase. The cDNA is then used as a template for exponential amplification using PCR. RT-PCR is currently the most sensitive method of RNA detection available. The use of RT-PCR for the detection of RNA transcript has revolutionalized the study of gene expression in the following important ways:
- Made it theoretically possible to detect the transcripts of practically any gene
- Enabled sample amplification and eliminated the need for abundant starting material that one faces when using northern blot analysis
- Provided tolerance for RNA degradation as long as the RNA spanning the primer is intact
Despite its major advantages, RT-PCR is not without drawbacks. The extreme sensitivity of the technique can be a double edged sword since even the slightest DNA contamination can lead to undesirable results. Additionally, planning and design of quantification studies can be technically challenging due to the existence of numerous sources of variation including template concentration and amplification efficiency.
Read more about this topic: Reverse Transcription Polymerase Chain Reaction
Famous quotes containing the word principles:
“My country is bleeding, my people are perishing around me. But I feel as a South Carolinian, I am bound to tell the North, go on! go on! Never falter, never abandon the principles which you have adopted.”
—Angelina Grimké (18051879)
“It is not impossible, of course, after such an administration as Roosevelts and after the change in method that I could not but adapt in view of my different way of looking at things, that questions should arise as to whether I should go back on the principles of the Roosevelt administration.... I have a government of limited power under a Constitution, and we have got to work out our problems on the basis of law. Now, if that is reactionary, then I am a reactionary.”
—William Howard Taft (18571930)
“Government ... thought [it] could transform the country through massive national programs, but often the programs did not work. Too often they only made things worse. In our rush to accomplish great deeds quickly, we trampled on sound principles of restraint and endangered the rights of individuals.”
—Gerald R. Ford (b. 1913)