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Buffer recipe for plasmid miniprep using spin columns

The cheapest but easy and effective way to get a plasmid miniprep kit is to buy spin columns for plasmid miniprep alone and make solutions with the following buffer recipe: Buffer P1: 50 mM Tris-HCl, 10 mM EDTA, pH 8.0 (25°C, the same below), 50 ug/ml RNase A Buffer P2: 0.2 M NaOH, 1% SDS […]


Plasmid miniprep protocols (spin columns)

Most of commercial plasmid miniprep kits rely on the spin column with silica membrane. The plasmid DNAs purified can be used for routine molecular biology applications such as DNA subcloning, PCR, in vitro transcription, DNA sequencing, and DNA transfection into mammalian cells. Protocols of plasmid miniprep using spin columns: – QIAprep Spin Miniprep Kit High-Yield […]


PCR product cloning protocols

Three mainstream strategies have been used to clone and subclone PCR-amplified DNA fragments. The traditional method is to digest the PCR products with restriction enzymes and ligate the digested DNA fragments with a digested vector. The second ligation strategy is called “TA cloning” based on the feature of ends of PCR products.  The modern cloning […]