Streamlined purification of plasmid DNA from prokaryotic cultures.

نویسندگان

  • Laura Pueschel
  • Hongshan Li
  • Matthew Hymes
چکیده

We describe the complete process of AcroPrep Advance Filter Plates for 96 plasmid preparations, starting from prokaryotic culture and ending with high purity DNA. Based on multi-well filtration for bacterial lysate clearance and DNA purification, this method creates a streamlined process for plasmid preparation. Filter plates containing silica-based media can easily be processed by vacuum filtration or centrifuge to yield appreciable quantities of plasmid DNA. Quantitative analyses determine the purified plasmid DNA is consistently of high quality with average OD(260/280;) ratios of 1.97. Overall, plasmid yields offer more pure DNA for downstream applications, such as sequencing and cloning. This streamlined method of using AcroPrep Advance Filter Plates allows for manual, semi-automated or fully-automated processing.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Cloning and Expression of Mycobacterium Tuberculosis ESAT-6 in Prokaryotic System

  The identification of a large number of antigens with potential for development of new tuberculosis vaccine has been accomplished in recent years. This study was designed for cloning and expression of ESAT-6 as a potent antigen of Mycobacterium tuberculosis. Selected gene (Rv3875) was amplified by PCR and product was ligated into expressing plasmid vector pQE30 and recombinant pQE30-ES plasmi...

متن کامل

بررسی مولکولی فعالیت پروتئولیتیکی پروتئین 3C در پاراکو ویروس انسانی تیپ یک

    Background & Aim: Human parechovirus is a genus of picornaviruses which includes a number of important viruses of clinical significance. Recent sequence analysis suggests that human parechovirus type 1(HPEV-1) is distinct from other picornaviruses and lacks the motives believed to be involved in the protease function of 2A. The aim of this study was to analyse proteolytic activity of 3C pro...

متن کامل

Prokaryotic expression, purification and immunogenicity analysis of CpsD protein from Streptococcus iniae

Streptococcus iniae is a major cause of serious bacterial infections in both fish and human beings. Capsular polysaccharide (CPS) of S. iniae is vital to evade phagocytic clearance of the host and serves as an important protective antigen of S. iniae infection in aquatic animals. The CpsD gene was determined to be highly conservative in capsule polysaccharide operon. Prokaryotic expression of t...

متن کامل

A REPORT OF RAPID AND EFFICIENT METHOD FOR MITOCHONDRIAL DNA

Based on the similarity of prokaryotic DNA to that of mitochondrial DNA, a rapid and efficient method for DNA purification from mitochondrial extracts obtained from different sources, was carried out based on the modification method described by Marmur (1983) which resulted in a high degree of purity (A260/A280=2). In addition we have applied the same technique for the first time to purify...

متن کامل

Prokaryotic expression, purification and immunogenicity analysis of CpsD protein from Streptococcus iniae

Streptococcus iniae is a major cause of serious bacterial infections in both fish and human beings. Capsular polysaccharide (CPS) of S. iniae is vital to evade phagocytic clearance of the host and serves as an important protective antigen of S. iniae infection in aquatic animals. The CpsD gene was determined to be highly conservative in capsule polysaccharide operon. Prokaryotic expression of t...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Journal of visualized experiments : JoVE

دوره 47  شماره 

صفحات  -

تاریخ انتشار 2011