Evaluation of silica resins for direct and efficient extraction of DNA from complex biological matrices in a miniaturized format.

نویسندگان

  • H Tian
  • A F Hühmer
  • J P Landers
چکیده

For DNA purification to be functionally integrated into the microchip for high-throughput DNA analysis, a miniaturized purification process must be developed that can be easily adapted to the microchip format. In this study, we evaluate the effectiveness of a variety of silica resins for miniaturized DNA purification and gauge the potential usefulness for on-chip solid-phase extraction. A micro-solid-phase extraction (muSPE) device containing only nanograms of silica resin is shown to be effective for the adsorption and desorption of DNA in the picogram-nanogram mass range. Fluorescence spectroscopy as well as capillary electrophoresis with laser-induced fluorescence detection is employed for the analysis of DNA recovered from solid-phase resins, while the polymerase chain reaction (PCR) is used to evaluate the amplifiable nature of the eluted DNA. We demonstrate that DNA can be directly recovered from white blood cells with an efficiency of roughly 70%, while greater than 80% of the protein is removed with a 500-nl bed volume muSPE process that takes less than 10 min. With a capacity in the range of 10-30 ng/mg of silica resin, we show that the DNA extracted from white blood cells, cultured cancer cells, and even whole blood on the low microliter scale is suitable for direct PCR amplification. The miniaturized format as well as rapid time frame for DNA extraction is compatible with the fast electrophoresis on microfabricated chips.

برای دانلود 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

An efficient and simple CTAB based method for total genomic DNA isolation from low amounts of aquatic plants leaves with a high level of secondary metabolites

An efficient DNA isolation protocol specifically modified to get pure quality DNA required for molecular studieshas been reported in this paper. Some aquatic plants (Potamogeton spp., Ceratophyllum demersum and Myriophyllum spicatum) were used for the study. The protocol developed will be useful in getting high and pure DNA. Instead of using the available DNA extraction kits, this protocol can ...

متن کامل

Evaluation of a modified salt-out method for DNA extraction from whole blood lymphocytes: A simple and economical method for gene polymorphism

Extraction of high-quality and-quantity DNA is a fundamental requirement for genetic research. It is very important to address the use of DNA extraction methods that are simple and cost-effective in gene polymorphism with large number of samples. This study was designed to investigate the optimal DNA extraction from lymphocytic cells by salt-out method. In this study, 200 blood samples of the t...

متن کامل

An Efficient Method For DNA Extraction From Paraffin Wax Embedded Tissues For PCR Amplification Of Human And Viral DNA

  Background and Objective: Formalin-fixed paraffin-embedded tissues are a valuable source of DNA for molecular studies. We designed and optimized an efficient procedure for DNA extraction from formalin-fixed paraffin embedded tissues. Materials and Methods: Seventy three blocks of cervical paraffin-embedded tissues were investigated. DNA was extracted using 45 minutes boiling in alkaline sol...

متن کامل

A simple and efficient DNA extraction protocol for old herbarium leaves of Bellevalia (Asparagaceae, Scilloideae)

High-quality DNA extraction plays an important role to make sharp bands in the gel electrophoresis and also produces clean chromatograms. Usually, DNA extract is delivered using the modified CTAB method but this method cannot obtain high-quality DNA for molecular analysis from old dried leaves of Bellevalia due to having different chemical compounds which inhibit to obtain a clear DNA extractio...

متن کامل

A Soft and Efficient Approach for Removal of Template from Mesoporous Silica using Benzene Sulfonamide

In this contribution, an effective and soft method for removal of template from nanochannels of mesoporous silica (MCM-41) is proposed. This method is based on chemically-modified solvent extraction which enhanced by means of an auxiliary organic compound, i.e. benzene sulfonamide. Template removal was performed in soft condition, i.e. in the presence of diluted sulfuric acid and at ambient tem...

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

برای دانلود 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

عنوان ژورنال:
  • Analytical biochemistry

دوره 283 2  شماره 

صفحات  -

تاریخ انتشار 2000