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Apparatus and method for electrophoresis

a technology of apparatus and electrophoresis, applied in the field of electrophoresis, can solve the problems of large buffer volume, environmental contamination of the system, and prior art electrophoresis systems are a potential source of contamination to the working environment, and achieve the effect of inhibiting the migration of ions

Inactive Publication Date: 2011-01-20
LIFE TECH ISRAEL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The solution effectively minimizes contamination, reduces the need for large buffer volumes, and maintains a stable pH during electrophoresis, enhancing the accuracy and speed of the separation process while preventing gas accumulation and ion interference with separated molecules.

Problems solved by technology

In addition, due to the use of buffers of low salt concentration, large volumes of buffer are required to maintain the electric field.
Prior art electrophoresis systems are a potential source of contamination to the working environment in which the tests are performed.
In addition, the environment is a potential source of contamination to the system, since PCR is an extremely sensitive method.
In fact, a single molecule of DNA product from one PCR (out of the trillions of molecules being produced) may interfere with the subsequent PCR such that it will produce incorrect results.
Conventional electrophoresis is also deficient in other respects, one being that it is time consuming.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

example 1

The ion source matrix 22 was prepared as follows:

A. a suspending gel of 3% agarose in 0.4X TAE buffer solution was prepared.

B. 0.6 grams of lead carbonate (PbCO3), prepared by bubbling CO2 into a solution of lead acetate following by filtration and wash by water of the precipitate, were suspended in 2 ml of the 3% agarose suspending gel of step A to obtain the ion source matrix 22.

C. A strip of lead was used as the cathode 26.

example 2

A. a suspending gel of 3% agarose in 0.4X TAE buffer solution was prepared.

B. 0.3 grams of silver chloride (AgCl) were suspended in 2 ml of the 3% agarose suspending gel of step A to obtain the ion source matrix 22.

C. A strip of aluminum was used as the cathode 26.

examples 3-10

Gel matrices containing several combinations of electrolyte solution were tested for use with a DNA sample (100 bp+1 kb ladder from Fermentas, Lithuaina) containing a tracking dye, such as bromophenol blue. The gel was run at 90-120V (currents=4-9 mAmps) until the bromophenol blue reached a distance of 5.7 cm from the wells. In these examples, an aluminum cathode was used in combination with a copper anode (Examples 3-8) or a silver anode (Examples 9 and 10). In the following examples, the electrolyte solution concentration was 100 mM, and the size of the cassette in length, width and thickness was 100 mm, 80 mm and 6.7 mm, respectively. Ions generated by the anode during electrophoresis procedure were inhibited when using the conditions describe hereinbelow. This phenomenon is likely caused by the formation of a salt complex.

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Abstract

Apparatus for conducting electrophoresis therein includes a chamber with a gel matrix. The chamber has a first sealed region and a second sealed region, and an anode within the first sealed region of the chamber and in contact with the gel matrix, and a cathode within the second sealed region and in contact with the gel matrix. At least one of the electrodes also provides ions for driving the electrophoresis. The apparatus further includes a matrix with at least one sparingly water-soluble salt.

Description

FIELD OF THE INVENTIONThe present invention relates to the field of electrophoresis and more particularly to apparatus for conducting electrophoresis tests therein.BACKGROUND OF THE INVENTIONA great deal of diagnostic procedures and laboratory research are carried out wherein DNA, RNA or proteins are mobilized and / or separated according to their physical and chemical properties via electrophoresis. This process is widely used and has may applications. For example, it is used to analyze DNA molecules according to their resultant size after being digested by restriction enzymes. It is also used to analyze the products of a polymerase chain reaction (PCR).Typically, electrophoresis separation is carried out in a separation matrix, such as a gel of agarose or acrylamide or a combination of the two. Usually, agarose gels are cast in open trays and form a slab whereas acrylamide gels are cast between two glass plates.In order to effect the electrophoretic process, two opposite ends of the...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N27/447B01D57/02
CPCB01D57/02G01N27/44721G01N27/44704
Inventor CABILLY, SHMUELYOGEV, URIMARGALIT, ILANA
Owner LIFE TECH ISRAEL
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