Plant tissue small sample rapid preparation method

A plant tissue and sample technology, applied in the field of rapid sample preparation, can solve problems such as low work efficiency, high extraction cost, loss of sample volume, etc., and achieve the effects of improving work efficiency, eliminating instrument operation, and saving test costs.

Inactive Publication Date: 2013-08-07
ZHENGZHOU TOBACCO RES INST OF CNTC
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Kits can be used to extract DNA from plant samples, but the extraction cost of each sample is relatively high, mainly for rapid sample preparation and DNA extraction of fresh plant samples with a small number of samples; another method is to use liquid nitrogen to freeze, and then magnetic beads High-frequency vibration is used to break the sample, and finally the DNA is extracted by the conventional CTAB or SDS method. There are many operating procedures, and the number of samples broken by the machine is small each time; the potassium content of plant tissue is mainly determined by flow analyzer and flame photometer. Determination method, but the sample preparation before the measurement on the machine usually uses a manual high-speed pulverizer to crush and dry the sample or use a mortar to grind the sample manually. The former requires a sufficient sampling amount for each sample, and the latter can grind a relatively small sample amount. samples, but the recovery rate is low, and there is a certain loss of sample volume. Both of them have large labor input and low work efficiency.

Method used

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  • Plant tissue small sample rapid preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0007] Example 1 Determination of Potassium Content in Tobacco Leaf

[0008] (1) Using the upper tobacco leaves of flue-cured tobacco NC297 as the material, 10 representative tobacco leaves were selected from the curing barn at different key stages of the tobacco leaf curing process, and punched with a 1 cm diameter hole punch on different parts of the tobacco leaves (7 Take 5 parts of the main leaf vein) punch holes to extract samples, put them directly into 2ml round-bottomed centrifuge tubes, all numbered on the centrifuge tubes, weigh the quality of the centrifuge tubes before loading the samples, and weigh the centrifuge tubes and tobacco leaves after loading the samples. . The sample prepared in the centrifuge tube was first opened and put in an oven at 105°C for 15 minutes, and then dried at 65°C. After drying, the quality of the centrifuge tube and the dried sample was weighed.

[0009] (2) Grind the drill bit of the electric drill into a nearly semicircular shape, a...

Embodiment 2

[0022] Example 2 Sample preparation for tobacco leaf DNA extraction

[0023] (1) Prepare a 2ml centrifuge tube, number it on the wall of the centrifuge tube with a pencil, cut the top young leaves of different tobacco materials with scissors, fold them up and place them in a 2 ml centrifuge tube, and put the centrifuge tube containing the sample in an ice box at low temperature save;

[0024] (2) Uncap the 2 ml centrifuge tube and throw it into a foam box filled with liquid nitrogen, then use a twister to take out the centrifuge tube that has been frozen quickly, and use an ordinary hand-held electrorotator with a round bottom rotor to rotate the sample at 2 ml centrifuge tube directly;

[0025] (3) Add 600 ul of CTAB solution preheated at 65°C (add 0.2% mercaptoethanol before use) to the centrifuge tube containing crushed tobacco leaves, and then put the centrifuge tube containing the CTAB-infiltrated sample into a 65°C water bath Water bath in the pot for about 1h;

[0...

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Abstract

The invention relates to a plant tissue small sample rapid preparation method which is used for carrying out component analysis or DNA identification of plant fresh samples. According to the invention, a plant fresh sample is directly added into a 2ml round-bottom centrifuge tube, and is subjected to a bake-drying treatment in a baking oven or a fast-drying treatment in liquid nitrogen; the sample is directly crushed in the centrifuge tube by using a handheld drill with a dome bit; an extraction liquid is added into the bake-dried sample tube for leaching, or an extraction liquid is added into the frozen sample tube for decomposing cells; a supernatant is obtained by separation in a high-speed centrifuge; and the supernatant is adopted as a component analysis or DNA identification sample. With the method provided by the invention, sample preparation work is flexible, instrument operation is eliminated, steps are simplified, certain test cost is saved, and working efficiency is improved. The method provided by the invention is suitable to be used in tobacco potassium nutrition and potassium high-efficiency genetic material potassium content determination works, and in sample preparation works before molecular biology research DNA extraction. The method facilitates laboratory sample analysis and testing works with small samples and large sample analysis amounts.

Description

technical field [0001] The invention relates to a rapid sample preparation method for analyzing and measuring plant tissues, in particular to a method for directly crushing fresh or dried plant samples in a centrifuge tube. Background technique [0002] Potassium (K + ) is the mineral nutrient element with the highest content in plants, also known as "quality element", which can increase crop yield, improve crop quality, and enhance crop resistance. Tobacco is a potassium-loving plant, and potassium is also one of the important quality indicators of flue-cured tobacco. High potassium content in tobacco leaves has a good effect on the appearance quality, sensory quality and smoking safety of tobacco leaves. At present, the potassium content of tobacco leaves in the Huanghuai tobacco area and some southwest tobacco areas in my country is relatively low. It is one of the economical and effective technical ways to increase the potassium content of tobacco leaves in my country t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/44
Inventor 高志强宋纪真周汉平王信民奚家勤郭建华
Owner ZHENGZHOU TOBACCO RES INST OF CNTC
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