Isolation of DNA and RNA Concepts

·Introduction:

Step 1: Sample Collection and Protection

Step 2: RNA Preparation

Step 3: Quantitation of Isolated RNA

Step 4: Storage of Isolated RNA

Step 1: Sample Collection and Protection

Finding the most appropriate method of cell or tissue disruption for your specific starting material is important for maximizing the yield and quality of your RNA preparation. During sample disruption for RNA isolation,

Samples stored at 4°C generate intact RNA, even after storage for a month.

Step 2: RNA Preparation

A number of RNA preparation technologies are widely available that can be classified into four general techniques:

organic extraction methods, spin basket formats, magnetic particle methods, and direct lysis methods.

While all can be used to prepare high-quality RNA suitable for a wide variety of analysis techniques, there are several factors to consider in selecting the right purification technology.

Direct Lysis Methods

Direct lysis methods perform sample preparation (not purification) by utilizing lysis buffer formulations that disrupt samples, stabilize nucleic acids, and are compatible with downstream analysis. Typically, a sample is mixed with lysis agent, incubated for some amount of time under specified conditions, and then used directly for downstream analysis. If desired, samples can often be purified from stabilized lysates. By eliminating the need to bind and elute from solid surfaces, direct lysis methods can avoid bias and recovery efficiency effects that may occur when using other purification methods.