Preparative and analytical purification of DNA from agarose.

نویسندگان

  • B Vogelstein
  • D Gillespie
چکیده

Two procedures were developed for removing DNA from agarose after electrophoretic separation of DNA fragments according to size. Both involve dissolving the DNA-containing agarose in NaI. The preparative technique uses binding of DNA to glass in the presence of NaI. The method is rapid and convenient, and DNA of all molecular weight ranges can be recovered in high yield and without degradation. The DNA is free of agarose and remains susceptible to digestion by restriction enzymes. The analytical technique uses selective precipitation of DNA with acetone and has been adapted to molecular hybridization scans of sequences in agarose gels. The sequence-monitoring system is quantitative, directly measuring the proportion of the probe complementary to a given DNA fragment and vice versa. It is especially suitable for analyzing restriction enzyme digests of DNA in mapping experiments.

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

Preparative separation of DNA--ethidium bromide complexes by zonal density gradient centrifugation.

Restriction enzyme fragmentation and recombinant DNA technology offer powerful tools with which to study gene structure, organization, and expression. Frequently in these studies the need arises to purify on a preparative scale a specific DNA molecule from a mixture. The two most commonly used procedures in such purification are isopycnic centrifugation in dye-salt gradients and gel electrophor...

متن کامل

Preparation of PCR products for DNA sequencing.

We demonstrate that routine PCR product analytical agarose gels can also serve as preparative gels for quick DNA template purification before sequencing. The band of interest is excised, placed into a Gel Nebulizer inside a Micropure separator and rapidly purified in a single centrifugation step. Gel-purified PCR product, suitable for manual and automated sequencing, is delivered within 10 min.

متن کامل

DNA Extraction from Beef Harboring Sarcocystis spp.: Comparison of Three Different Analytical Methods

Background: DNA extraction is one the most important steps for molecular analysis of food-borne pathogens. In this research, three methods of DNA extractions from beef harboring Sarcocystis spp. were compared for the quality, quantity, safety, as well as cost-effectiveness. Methods: About 100 mg intersostal and diaphragm were collected from 10 slaughtered cattle. After ensuring their contami...

متن کامل

Section Ii Resolution and Recovery of Large Dna Fragments

This section describes the application of agarose gel electrophoresis to both analytical and preparative separation of DNA fragments. Standard agarose gels separate DNA fragments from ∼0.5 to 25 kb, whereas pulsed-field agarose gels resolve molecules from ∼10 to >2000 kb. Descriptions of standard and pulsed-field agarose gel electrophoresis as well as parameters affecting resolution of large DN...

متن کامل

Purification of Bacillus subtilis RNA polymerase with heparin-agarose. In vitro transcription of phi 29 DNA.

We have devised a new procedure for the purification of highly active preparations of Bacillus subtilis RNA polymerase holoenzyme. A column of heparin-agarose A-15m is used to rapidly and quantitatively adsorb RNA polymerase from the initial crude extract fraction. This affinity procedure obviates the necessity of including nucleic acid precipitation or partitioning steps and allows for rapid s...

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:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 76 2  شماره 

صفحات  -

تاریخ انتشار 1979