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In the A/T site, the A-site half resides in the small ribosomal subunit where the mRNA decoding site is located. The mRNA decoding site is where the mRNA codon is read out during translation. The T-site half resides mainly on the large ribosomal subunit where EF-Tu or eEF-1 interacts with the ribosome.
Competitive RT-PCR technique is used for absolute quantification. It involves the use of a synthetic “competitor” RNA that can be distinguished from the target RNA by a small difference in size or sequence. It is important for the design of the synthetic RNA be identical in sequence but slightly shorter than the target RNA for accurate results.
Protein synthesis takes place in cytosolic ribosomes, mitochondria (mitoribosomes), and in plants, the plastids (chloroplast ribosomes). Each of these compartments requires a complete set of functional tRNAs to carry out protein synthesis.
RNA-Seq methodology has constantly improved, primarily through the development of DNA sequencing technologies to increase throughput, accuracy, and read length. [61] Since the first descriptions in 2006 and 2008, [40] [62] RNA-Seq has been rapidly adopted and overtook microarrays as the dominant transcriptomics technique in 2015. [63]
In order to robustly detect and quantify gene expression from small amounts of RNA, amplification of the gene transcript is necessary. The polymerase chain reaction (PCR) is a common method for amplifying DNA; for RNA-based PCR the RNA sample is first reverse-transcribed to complementary DNA (cDNA) with reverse transcriptase.
Transcription-mediated amplification has several advantages compared to other amplification methods including: TMA is isothermal, meaning it is performed at constant temperature. As such, a water bath or heat block can be used instead of a thermal cycler. TMA produces RNA amplicon rather than DNA amplicon.
Student's t-test, Analysis of variance, Mann–Whitney U test: Repeated measures design: A research design that involves multiple measures of the same variable taken on the same or matched subjects either under different conditions or over two or more time periods. [1] Paired t-test, Wilcoxon signed-rank test
The presence of this functional group causes the helix to mostly take the A-form geometry, [11] although in single strand dinucleotide contexts, RNA can rarely also adopt the B-form most commonly observed in DNA. [12] The A-form geometry results in a very deep and narrow major groove and a shallow and wide minor groove. [13]