Helge Bastian

DNA Extraction Using Anion-Exchange Chromatography and Silica-Gel Based Membranes

Reliable and efficient methods for the isolation of nucleic acids are an absolute prerequisite for their analysis by molecular biological methods. Techniques, such as AFLP, RFLP, PCR, RT-PCR, library construction, and many other downstream applications, depend upon pure and intact DNA or RNA, since residual impurities in the nucleic acid preparation can inhibit enzymatic reactions, electrophoresis and quantification. Methods for the isolation of nucleic acids from complex starting materials, like animal and plant tissues, usually involve a lysis step, to release the cell contents, followed by separation of the nucleic acid from other cellular components. Conventional isolation protocols use CsCl2 gradients (1, 2), or extraction with phenol and/or chloroform (3,4).

Isolation of Total RNA Using Silica-Gel Based Membranes

Procedures are given for both plant and animal tissues. They differ in the preparation of the lysate, after which the steps are the same for both plant and animal samples.