A tool for high-throughput plant (transfer) gene detection---plant material direct PCR technology

A tool for high-throughput plant (transfer) gene detection---plant material direct PCR technology
Abstract: Beijing Baitek Biotechnology Co., Ltd. and the General Plant Total RNA Extraction Kit (RP3301) have successfully solved the problems of various difficult RNAs, and recently introduced the latest products for the extraction and amplification of high-throughput plant DNA. ——Plant Direct PCR Kit (PR7501), which does not require liquid nitrogen and grinding 杵 grinding, nor does it require steel ball crushing, and does not rely on any special equipment, and the method is convenient and quick.
It is well known that in the genetic mapping of forward genetics, plant quality and variety identification, or detection of plant active ingredients, and transgenic detection, DNA extraction from high-throughput plants is required, followed by PCR. Especially in gene map cloning, it is necessary to construct a large population (several thousands, more than tens of thousands), then extract DNA, PCR method to screen the linkage markers of traits (SSR, RAPD, etc.), and finally sequence clone gene. In the conventional method, in the DNA extraction, the sample is ground with liquid nitrogen, then extracted with chloroform, and finally PCR. The extraction of thousands of strains of DNA takes several weeks. Some companies have made improvements and developed a relatively simple method. Extraction with a 96-well adsorption plate takes about one hour to complete the experiment. Some companies also add steel balls or other small metals to the 96-well deep-well plate. The beads help to break, the supernatant is directly lysed and the supernatant is taken for PCR, which can shorten the time to half an hour, but requires a special oscillator, and the use cost of the steel ball is high; some companies provide small grinding burrs without the use of steel balls. After the PCR tube is artificially ground, the supernatant is lysed and subjected to PCR, which obviously consumes a lot of labor time, and is also easy to contaminate each other.
The above methods are obviously not ideal for large-scale gene or transgenic detection, not only for a long time but also for pollution! Many well-known companies at home and abroad have invested in improving the research and development of new technologies! Beijing Baitek Biotechnology Co., Ltd. and the General Plant Total RNA Extraction Kit (RP3301) successfully solved the problem of various difficult RNAs, and recently launched the latest products for the extraction and amplification of high-throughput plant DNA. Plant Direct PCR Kit (PR7501), which does not require liquid nitrogen and grinding, does not require steel ball crushing, and does not require any special equipment, and is convenient and quick. Simply take 0.5-0.7 cm of plant leaves into solution L, incubate at 95 °C for 10 minutes, without liquid nitrogen grinding, then add an equal volume of solution I, you can directly carry out PCR, convenient and fast, is the research work in the field of plants One of the best choices. The characteristics of the plant direct PCR kit are:
Ø Plant genomic DNA can be extracted in one step in 12 minutes .
Ø No phenol / chloroform extraction is required.
Ø need not be frozen in liquid nitrogen plant tissue, mechanical treatment, or purification of the DNA was precipitated.
Ø containing PCR MIX, 4ul can take direct PCR after lysis.
Ø No need to load buffer and dye.
Ø The extract can be stored for 6 weeks at 4 °C .
Example 1 :
1. Cut 1-2 pieces of 0.5-0.7 cm rice leaves into a 1.5 ml centrifuge tube and add 100 μl of solution L to ensure that the leaves are immersed in the solution.
2. Incubate at 95 ° C for 10 minutes, at which time the DNA has been released but the leaves are not significantly digested.
3. Add 100 μl of solution I. Vortex mix. Direct PCR.
4. Preparation of PCR reaction system:
PCR system
volume
Extract
4μl
2×PCR MIX
10μl
Upstream primer
1μl
Downstream primer
1μl
water
4 μl
total capacity
20μl
5. Perform the following reactions on the PCR machine:
94 ° C 3 min
94°C 0.5-1min
50-65 C 0.5-1min 30-35 cycles
72 ° C 0.5-2 min
72 ° C 10 min
6, 8% polyacrylamide gel electrophoresis, silver staining.

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