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Kit and method for extracting DNAs in agarose gel by using magnetic beads

An agarose gel and kit technology, applied in the field of molecular biology, can solve the problems of increased time-consuming purification, increased workload of experimental personnel, increased human error, etc., and achieves high DNA extraction recovery rate, improved work efficiency, The effect of convenient automatic dosing

Inactive Publication Date: 2017-07-04
SUZHOU ENRICHING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method of using gold magnetic particles to purify and recover DNA in agarose gel is easy to operate, but the disadvantage is that the entire operation needs to be carried out in ep tubes. When the amount of samples increases, the purification time will increase and the workload of the experimenters will increase. significantly increased, and the human error also increased during the experiment

Method used

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  • Kit and method for extracting DNAs in agarose gel by using magnetic beads
  • Kit and method for extracting DNAs in agarose gel by using magnetic beads

Examples

Experimental program
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Effect test

Embodiment 1

[0036] Embodiment 1: Magnetic bead method agarose gel DNA extraction kit:

[0037] Magnetic Beads Agarose Gel DNA Extraction Kit includes:

[0038] Sol-binding solution: a mixed solution of pH 5.2, 6M NaI, and 0.03M sodium acetate. Specific preparation method: Weigh 90g of sodium iodide and 0.25g of sodium acetate, add enough distilled water to fully dissolve, continue to add distilled water to set the volume to 100mL, and adjust the pH value to 5.2 with glacial acetic acid.

[0039] Magnetic bead dispersion: magnetic bead suspension with a concentration of 20 μg / uL, the inner core of the magnetic bead is ferric oxide (Fe 3 o 4 ), the surface is coated with SiO 2 layer. Specific preparation method: Weigh 2g of magnetic beads, add to 100mL of distilled water, and disperse evenly by ultrasonic.

[0040] Cleaning solution I: 40mM Tris-HCl, 10mM EDTA and 50% (v / v) isopropanol aqueous solution. Specific preparation method: Weigh 0.63g Tris-HCl and 0.29g EDTA, add enough 50% (...

Embodiment 2

[0063] Example 2: High-throughput DNA extraction from agarose gel after electrophoresis of 96 copies of 600bp DNA fragments.

[0064] The kit described in Example 1 was used for extraction.

[0065] Step 1: Sample Preparation

[0066] Take 96 copies of 50ng / uL X 6uL 600bp DNA fragments, and use 1% agarose gel for electrophoresis in TAE buffer. Cut out the gel, keep the weight of the gel block less than 400mg, put the gel block into a clean 96-well circular well plate, and parallel 96 samples in total.

[0067] Step 2: Sol

[0068] Add 500uL of binding solution to 96 sample wells with a multi-channel automatic liquid adder. Place the 96-well plate in a 65 °C water bath for 10 min.

[0069] Step 3: Binding of magnetic beads to DNA

[0070] Take the 96-well plate out of the water bath, and add 50uL magnetic bead dispersion to the 96 sample wells with an 8-channel automatic liquid dispenser. After addition, the 96-well plate was placed on a vortex mixer and shaken at 900 rpm...

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Abstract

The invention discloses a kit for extracting DNAs in agarose gel by using magnetic beads and a method for high-flux rapid extraction of DNAs in agarose gel. The kit comprises a sol binding solution, a magnetic bead dispersion solution, a rinsing solution I, a rinsing solution II and an eluting solution. According to the invention, the kit is creatively used together with a 96-well plate, a magnetic plate frame for the 96-well plate, a 96-site bar magnet claw and a vortex mixer for completion of extraction, so extraction of DNAs from agarose gel can be directly carried out in the 96-well plate; in the process of an experiment, feeding of each test buffer solution can be rapidly carried out by using a multichannel automatic liquid feeding device; and one 96-well plate can be used for DNA extraction of 96 samples. With the kit and the high-flux method, when an individual experimenter operates six 96-well plates at one time, DNA extraction of 576 samples can be finished within about 70 to 80 min.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a magnetic bead method agarose gel DNA extraction kit and a method for rapidly extracting agarose gel DNA with high throughput. Background technique [0002] In molecular biology experiments, recovering and purifying DNA fragments from agarose gel after electrophoresis is one of the most commonly used techniques and has a history of nearly 40 years. Early methods for recovering DNA from agarose gel included low-melting point agarose gel method, dialysis charged elution method, DEAE cellulose membrane disc method, etc., all of which required organic reagents such as phenols and chloroform for extraction Extraction, and then precipitation with ethanol, the operation process often takes a long time, and it takes nearly 2 hours for one sample. This kind of method is currently rarely used. Later, the development of column DNA recovery kits greatly improved the convenience...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 吴巧尚春庆黄娱琴雷波赵青玲曲峰
Owner SUZHOU ENRICHING BIOTECH CO LTD
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