Is RNA extraction hard?
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| Papers (10) | Insight |
|---|---|
67 Citations | So it is extremely difficult for RNA extraction from its various tissues, which prevents following molecular operations and expression-level studies like microarray and RT-PCR. |
45 Citations | RNases may be difficult to eliminate, although with care and appropriate countermeasures, reproducible extraction of high-quality RNA from important biological samples should be attainable. |
23 Citations | With our optimized RNA extraction protocol, there is no difficulty in extracting great amounts of RNA with high quality. |
| Here we report a method for RNA extraction that takes about 40 min to complete ten samples, and is not more laborious than current commercial RNA extraction kits. | |
| This emerging RNA extraction method is direct, rapid, and does not require the use of specialized equipment, thus offering advantageous application for laboratories with limited resources. | |
18 Citations | In practice, this means that a selective extraction process is required to remove all the unwanted cellular material in a manner that minimizes degradation of the RNA by hydrolysis or ribonuclease activity. |
16 Citations | This study investigated the cause of this degradation and identifies an alternative procedure that enables extraction of intact high-quality RNA. |
23 Citations | The extraction provides high yield and the extracted RNA is undegraded. |
| Researchers should optimise these methods for their specific application and keep in mind that “total RNA” extraction methods do not isolate all types of RNA equally. | |
61 Citations | This paper presents a novel, highly efficient and simple method for low-molecular weight RNA extraction using electricity only. |
Related Questions
Amount of starting material for RNA extraction for RNA sequencing4 answersRNA sequencing (RNA-seq) can be performed using low amounts of starting material, ranging from 10 pg to 10 ng of total RNA or 1-1000 intact cells. For gene expression analysis from seed tissues, high-quality RNA can be extracted using only 2 to 3 mg of starting tissue. A new RNA-Seq library preparation technique has been developed that allows representative, strand-specific RNA-Seq libraries to be generated from small amounts of starting material in a fast and cost-effective manner. RNA can also be extracted from dried blood spots (DBS) for transcriptome studies, providing an option when field conditions make it difficult to collect, store, or transport whole blood for RNA studies. Additionally, a miniSAGE technique has been established for determining gene expression patterns using as little as 1 μg of RNA as starting material.
Why is RNA extracted and quantified?4 answersRNA is extracted and quantified for several reasons. Accurate quantification of RNA is important for downstream RNA analysis techniques. It helps in determining the concentration of RNA in a sample, which is necessary for performing optimal RNA analysis. Different methods, such as UV spectrophotometric and fluorescent dye-based methods, are used for quantifying RNA depending on the purity and concentration of the sample. RNA extraction is also necessary for cell characterization and studying gene expression. In the context of infection diagnosis, measuring RNA expression profiles can provide a specific systemic inflammatory response to infection, which can be used as a molecular diagnostic approach. Additionally, accurate RNA quantification is important for quality control in RNA synthesis and extraction processes. Overall, RNA extraction and quantification are essential steps in various research and diagnostic applications.
How to extract rna from blood sample using trizol method step by step?4 answersTo extract RNA from a blood sample using the Trizol method, the following steps can be followed. First, collect the blood using a negative pressure blood sampling tube filled with an anticoagulant and Trizol agent. Store the blood at room temperature for 30 days. Next, add Trizol reagent to the pretreated sample in an RNase-free centrifuge tube and mix. Extract chloroform and add isopropanol to obtain a white sediment. Wash and centrifuge the sediment, then discard the supernatant. Finally, add diethyl pyrocarbonate to dissolve the sediment and obtain a purified RNA sample. Another method involves using an integrated centrifugo-pneumatic micro-homogenizer, which efficiently lyses whole blood and yields the aqueous phase with the total RNA content. These methods provide high-quality RNA extraction from blood samples.
Does RNA come in aqueous phase during DNA isolation?5 answersRNA is concentrated in the aqueous phase during DNA isolation.
What is the principle of magnet-based RNA extraction?2 answersMagnet-based RNA extraction involves the use of magnetic beads to efficiently extract RNA from a sample. The process typically includes the following steps: (1) The sample, such as thallus or a viral sample, is collected and placed in a centrifuge tube. (2) Buffer solutions containing lysozyme and other reagents are added to the tube to resuspend the sample and release the RNA. (3) The mixture is then subjected to vortex oscillation and centrifugation to separate the insoluble precipitate and obtain the supernatant containing the RNA. (4) Buffer solutions and magnetic bead solutions are added to the tube, and the mixture is uniformly mixed. The magnetic beads, which are coated with silica or other materials, bind to the RNA, allowing for its concentration and extraction. This method offers advantages such as increased extraction efficiency, simplicity, and quick operation.
Plant RNA extraction how to get rid of protein contamination?5 answersTo get rid of protein contamination during plant RNA extraction, several methods can be used. One approach is to use a homemade AGPC solution, which involves a phase separation process to isolate RNA while leaving proteins in the interphase and organic phase. Another method involves crushing plant samples in batches using liquid nitrogen and steel balls, which helps to avoid degradation of RNA and cross-infection. Additionally, using TRizol, chloroform, and isopropanol in the extraction process can help remove protein contamination and obtain high-quality RNA. Another extraction method involves grinding plant materials with liquid nitrogen, performing water-saturated phenol and chloroform extraction, and using isopropanol precipitation to obtain total RNA, which effectively avoids the influence of LiCl residues on downstream experiments. Finally, a method using sorbitol, polyvinylpyrrolidone, bovine serum albumin, polyethylene glycol, and spermidine can be used to precipitate total nucleic acid while removing most secondary metabolites, followed by the removal of proteins using chloroform and precipitation of RNA with ethanol.