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When using a path length that is shorter than 10mm, the resultant OD will be reduced by a factor of 10/path length. Using the example above with a 3 mm path length, the OD for the 100 μg/mL sample would be reduced to 0.6. To normalize the concentration to a 10mm equivalent, the following is done: 0.6 OD X (10/3) * 50 μg/mL=100 μg/mL
The Qubit fluorometer method is to use fluorescent dyes to determine the concentration of either nucleic acids or proteins in a sample. Specialized fluorescent dyes bind specifically to the substances of interest. A spectrophotometer is used in this method to measure the natural absorbance of light at 260 nm (for DNA and RNA) or 280 nm (for ...
RNA-Skim RNA-Skim: a rapid method for RNA-Seq quantification at transcript-level. rSeq rSeq is a set of tools for RNA-Seq data analysis. It consists of programs that deal with many aspects of RNA-Seq data analysis, such as read quality assessment, reference sequence generation, sequence mapping, gene and isoform expressions (RPKMs) estimation, etc.
2 Quantification. 3 Synthesis. 4 Kinetics. 5 Gene function. 6 Other. 7 See also. 8 References. 9 External links. Toggle the table of contents. ... DNA and RNA ...
The purified RNA is analysed for quality (by capillary electrophoresis) and quantity (for example, by using a NanoDrop or NanoPhotometer spectrometer). If the material is of acceptable quality and sufficient quantity is present (e.g., >1μg, although the required amount varies by microarray platform), the experiment can proceed.
Gene expression and RNA quantification studies have benefited from the increased precision and absolute quantification of dPCR. [91] RNA quantification can be accomplished via RT-PCR, wherein RNA is reverse-transcribed into cDNA in the partitioned reaction itself, and the number of RNA molecules originating from each transcript (or allelic ...
[25] [60] Theoretically, there is no upper limit of quantification in RNA-Seq, and background noise is very low for 100 bp reads in non-repetitive regions. [ 10 ] RNA-Seq may be used to identify genes within a genome , or identify which genes are active at a particular point in time, and read counts can be used to accurately model the relative ...
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.
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