High-throughput plasmid mini preparations facilitated by micro-mixing.
نویسندگان
چکیده
We have developed a reliable high-throughput plasmid isolation system using a 96-well plate format. This system combines a novel glass bead micro-mixing method with modified alkaline lysis and Sephacryl S-500 DNA purification procedures. Mechanical forces generated by vortexing glass beads inside each well of the 96-well plates ensure that the bacterial pellets are homogeneously resuspended, the cells are completely lyzed, and the resulting bacterial lysates are thoroughly mixed with the potassium acetate solution. The vortexing speed and duration for glass bead mixing have been standardized to facilitate plasmid DNA yields without significant adjustments.
منابع مشابه
Transferable neuronal mini-cultures to accelerate screening in primary and induced pluripotent stem cell-derived neurons
The effort and cost of obtaining neurons for large-scale screens has limited drug discovery in neuroscience. To overcome these obstacles, we fabricated arrays of releasable polystyrene micro-rafts to generate thousands of uniform, mobile neuron mini-cultures. These mini-cultures sustain synaptically-active neurons which can be easily transferred, thus increasing screening throughput by >30-fold...
متن کاملExperimental and Computational Study on Hydrodynamic of a Downscaled Mini Vessel USP Dissolution Test Apparatus II
Although not listed on the United States Pharmacopeia (USP), like standard USP 2, small volume USP 2 dissolution apparatus has gained a great deal of attention, especially for cases where small amount of drug product is available for testing in research and design step or evaluations are to be made on a tablet containing trace amounts of the active pharmaceutical ingredient. In this work, first...
متن کاملNumerical Investigation of Fluid Mixing in a Micro-Channel Mixer with Two Rotating Stirrers by Using the Incompressible SPH Method
Fluid mixing is a crucial and challenging process for microfluidic systems, which are widely used in biochemical processes. Because of their fast performance, active micromixers that use stirrer blades are considered for biological applications. In the present study, by using a robust and convenient Incompressible Smoothed Particle Hydrodynamics (ISPH) method, miscible mix...
متن کاملHigh-throughput droplet PCR.
The polymerase chain reaction has facilitated the ready analysis of nucleic acids. A next challenge requires the development of means to unravel the complexity of heterogeneous tissues. This has presented the task of producing massively parallelized quantitative nucleic acid data from the cellular constituents of tissues. The production of aqueous droplets in a two phase flow is shown to be rea...
متن کاملNumerical Study on Low Reynolds Mixing ofT-Shaped Micro-Mixers with Obstacles
Micromixers are one of the most crucial components of Lab-On-a-Chip devices with the intention of mixing and dispersion of reagents like small molecules and particles. The challenge facing micromixers is typically insufficient mixing efficiency in basic designs, which results in longer microchannels. Therefore, it is desirable to increase mixing efficiency, in order to decrease mixing length, w...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Nucleic acids research
دوره 24 24 شماره
صفحات -
تاریخ انتشار 1996