Hvorfor nedbrydes RNA lettere end DNA under nukleinsyreekstraktion?

Grunden til, at RNA (ribonukleinsyre) nedbrydes lettere end DNA (deoxyribonukleinsyre) during udvinding is mainly related to their different chemical structures and improper operation during extraction.

jeg. Differences in chemical structure:

1. RNA is a single-stranded structure, which is more unstable than the double-stranded structure of DNA.

2. Hydroxyl at the 2′ position: The ribose in RNA molecules contains a hydroxyl group (-Åh) at the 2′ position, while the deoxyribose in DNA has a hydrogen atom at this position. This extra hydroxyl group makes the RNA molecule more unstable because it can promote spontaneous hydrolysis reactions, resulting in RNA chain breaks.

II. The influence of RNAse in the extraction system:

1. Endogenous RNAse: It exists in cells and can continue to degrade RNA even after cell lysis.

2. Exogenous RNAse: It is widely present in the environment, such as air, skin, and laboratory supplies (gloves, test tubes, osv.).

III. Improper experimental operation:

1. The reagents and instruments used have not been treated with DEPC (diethylpyrocarbonate) and cannot effectively inactivate RNA enzymes.

2. The inhibitor is ineffective, or the dosage is insufficient, and the activity of RNA enzymes cannot be effectively inhibited.

3. More experimental samples or higher homogenization temperatures lead to increased RNA enzyme activity.

4. Changes in pH and temperature, especially under alkaline conditions, can easily cause RNA bases to be deaminated, resulting in chain breaks; high temperatures can also cause partial or complete degradation of RNA molecules.

IV. What should be done to reduce RNA degradation？

In order to reduce RNA degradation during extraction, a series of measures need to be taken, including using RNAse-free laboratory supplies, operating at low temperatures, using RNA stabilizers, using finished kits, osv. These methods effectively reduce the risk of RNA degradation and ensure that high-quality RNA samples are obtained for downstream analysis.

Leverandørintroduktion

Shanghai Lingjun Biotechnology Co., Ltd.blev etableret i 2016 som er en professionel producent af biomagnetiske materialer og nukleinsyreekstraktionsreagenser.

Vi har stor erfaring med udvinding og oprensning af nukleinsyre, proteinoprensning, celleadskillelse, kemiluminescens og andre tekniske områder.

Vores produkter er meget udbredt inden for mange områder, såsom medicinsk test, genetisk testning, universitetsforskning, genetisk avl, og så videre. Vi leverer ikke kun produkter, men kan også påtage os OEM, ODM, og andre behov. Hvis du har relaterede behov, er du velkommen til at kontakte os på sales01@lingjunbio.com.