Simple and rapid preparation of plasmid template by a filtration method using microtiter filter plates.

نویسندگان

  • M Itoh
  • P Carninci
  • S Nagaoka
  • N Sasaki
  • Y Okazaki
  • T Ohsumi
  • M Muramatsu
  • Y Hayashizaki
چکیده

We developed a new simple high-throughput plasmid DNA extraction procedure, based on a modified alkaline lysis method, using only one 96-well microtiter glassfilter plate. In this method, cell harvesting, lysis by alkaline and plasmid purification are performed on only one microtiter glassfilter plate. After washing out RNAs or other contaminants, plasmid DNA is eluted by low-ion strength solution, although precipitated chromosomal DNA is not eluted. The plasmid prepared by this method can be applied to sequencing reactions or restriction enzyme cleavage.

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

روش سریع و مؤثر ترانسفورم‌سازی باکتری اشریشیاکلی

  Background & Objective: Transformation of plasmid DNA into bacterial competent cells is a key technique for molecular cloning. Transformation can be achieved using either chemical or physical methods, e.g., electroporation. The rate of success in these methods depends on experience and attention to method’s details. Therefore, the higher the efficiency and quality of a transformation method, ...

متن کامل

Streamlined purification of plasmid DNA from prokaryotic cultures.

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 filtr...

متن کامل

High-speed plasmid isolation using 96-well, size-exclusion filter plates.

The accelerating pace of genomics analysis has necessitated the abbreviation of DNA sample preparation protocols. We have developed a size-exclusion-based system for the rapid isolation ofplasmid DNA in a 96-well microplate format. This high-speed protocol employs a modified alkaline lysis methodfor the preparation of the bacterial lysate, followed by three short vacuum filtration steps. Unlike...

متن کامل

An automated method for DNA preparation from thousands of YAC clones.

We describe an automated method for the preparation of yeast genomic DNA capable of preparing thousands of DNAs in parallel from a YAC library. Briefly, the protocol involves four steps: (1) Yeast clones are grown in the wells of 96-well microtiter plates with filter (rather than plastic) well-bottoms, which are embedded in solid growth media; (2) These yeast cultures are resuspended and their ...

متن کامل

Investigating the Influence of Filter Uniformity Coefficient and Effective Pore Size on Critical Hydraulic Gradient and Maximum Erosion of Dispersive and Non-dispersive Samples

Filter is one of the main components of embankment dams. By a simple but effective performance, filter protects the dam against erosion and soil scouring in impervious core caused by leakage (piping) and makes it safe. Interaction between filter and erodible base soil is a complex phenomenon which is dependent upon several factors, and has challenged researchers for better understanding the fil...

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:
  • Nucleic acids research

دوره 25 6  شماره 

صفحات  -

تاریخ انتشار 1997