Veiledning til effektiv RNA-ekstraksjon: Fra prinsipper til praksis, sikre RNA-isolering av høy kvalitet

Introduction of RNA Extraction

RNA extraction is a basic step in molecular biology and gene expression research. Dens formål er å isolere intakt, ikke-nedbrutt RNA (ribonukleinsyre) from cell or tissue samples. RNA participates in a variety of biological processes, such as transcription of genetic information and protein synthesis in cells. Derfor, obtaining high-quality RNA is crucial for subsequent molecular biological analysis, such as RT-PCR, Northern Blot, RNA sequencing, osv.

Principle of RNA Extraction

Denaturants break cells or tissues, and then RNA is extracted by organic solvents such as chloroform and then precipitated, washed, dried, and finally dissolved.

Imidlertid, since RNA enzymes are everywhere and may degrade RNA at any time, there are many places to pay attention to in the experiment. Any negligence will fail.

Avoid detours in RNA extraction

How to choose the most suitable RNA extraction reagent for different materials.

The numerous existing RNA isolation methods may make it difficult to choose. For tiden, the simplest and safest method is column separation, such as RNApure, which is popular because of its simple operation, short time and high purity. RNApure does not require DNase to digest DNA, which saves time and avoids RNA degradation, thereby increasing the yield; compared with non-column separation (such as TRIzol), it removes proteins and other impurities cleanly, improves purity, and is very suitable for cells, vev, and general plants. The extraction of plant RNA is more difficult to choose. Plant RNA extraction is affected by the content of phenol, polysakkarider, protein impurities, og sekundære metabolitter. Generally, the purity of TRIzol extraction is not high, and many plants cannot be extracted with TRIzol.

At this time, you can choose polysaccharide and polyphenol plant total RNA extraction kit (daffodils, peppers, carrots, corn, lilies, wheat, tomatoes, cauliflower, rapeseed, osv.) and universal plant RNA extraction kit (suitable for most plant extractions, including various Chinese herbal medicines, plant seeds, apples, grapes, jordbær, bananas, longans, litchi, lawn plants, pine trees, fir trees, birch trees, purple pine cones, coleus, poinsettia, oleander, fig, violet, rose, dahlia, morning glory, osv.); It is worth mentioning that the extraction of RNA from blood (inkludert serum, plasma, cerebrospinal fluid, various swabs or other samples with high liquid content) using TRIzol and red blood cell lysis methods are not effective because red blood cell lysis buffer does not contain RNase inhibitors, and RNA is easily degraded in this process. It is recommended to use TRIpure LS (RP1101) or a blood total RNA extraction kit (RP4001). This method is suitable for RNA extraction in expression profile chip experiments and has become a nationally recommended method.

How to store extracted RNA?

If it is only for short-term storage, the resuspended RNA should be placed at -20°C; if it is for long-term storage, it should be placed at -80°C. When storing RNA, a small amount of RNase inhibitor (RP5601) can be added to avoid RNA degradation, and RNA can be directly used for downstream experiments. If RNA is to be stored for a long time, RNAlong can be added. We recommend that the RNA solution be divided into several tubes. This will avoid repeated freezing and thawing to damage the RNA and prevent accidental RNase contamination.

Leverandør

Shanghai Lingjun Biotechnology Co., Ltd. ble etablert i 2016 som er en profesjonell produsent av biomagnetiske materialer og nukleinsyreekstraksjonsreagenser.

Vi har rik erfaring innen utvinning og rensing av nukleinsyre, proteinrensing, celleseparasjon, kjemiluminescens og andre tekniske felt.

Våre produkter er mye brukt på mange felt, som medisinsk testing, genetisk testing, universitetsforskning, genetisk avl, og så videre. Vi leverer ikke bare produkter, men kan også påta oss OEM, ODM, og andre behov. Hvis du har et relatert behov, ta gjerne kontakt med oss ​​på sales01@lingjunbio.com.