Method for extracting genome DNA of fungus
A genome and fungal technology, applied in the field of extracting fungal genomic DNA, can solve the problems of low extraction rate, time-consuming and laborious, etc., and achieve the effect of simple extraction process and low cost
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0068] Example 1: Genomic DNA of filamentous fungal hyphae is extracted
[0069] Genomic DNA was extracted from endophytic fungi isolated from corn roots and leaves. The samples were: "1": T4R6A, "2": T4R6B, "3": T4R6B1, "4": T1R2A, "5": T4R5A , "6": T4R5B, "7": T4R2B, "8": T4R2A.
[0070] Extraction steps: (1) Under the ultra-clean workbench, use a sterile skipping needle to pick a small amount of mycelium cultured in solid medium and put it in a PCR tube prepared in advance with ES solution; (2) PCR Put the tube into the PCR machine, and then incubate at 95°C for 10 minutes; (3) After taking it out, add BSA solution 3 times the volume of the ES solution, and the extraction is complete; (4) Verify the DNA extraction effect, and perform PCR amplification on the LSU fragment , the amplification result is as figure 1 : The original DNA of 8 samples was diluted 10-fold and 50-fold and then amplified. The results of electrophoresis showed that except for the weak band in the 10-...
Embodiment 2
[0071] Embodiment 2: the genomic DNA of mycorrhizal sample is extracted
[0072] The samples were obtained from synthetic seedlings of truffles and oak tree mycorrhizae in a laboratory greenhouse. The samples were chestnut × rufous mushroom, Mongolian oak × russia, hazelnut × russia, and Korean pine × russia.
[0073] Extraction steps: (1) Take a small amount of root samples, rinse with water to remove the culture medium, put them into a clean petri dish, add a small amount of water, observe under a dissecting microscope, pick 3 root tips and put them into the prepared ES solution (2) Put the PCR tube into the PCR instrument and incubate at 95°C for 10 min; (3) After taking it out, add BSA solution 3 times the volume of the ES solution. Save for later use; (4) Verify the DNA extraction effect, and perform PCR amplification on the ITS fragment, and the amplification results are as follows: figure 2 : "1" is the synthetic mycorrhizal sample of chestnut × milk mushroom, "2" is...
Embodiment 3
[0074] Example 3: Extraction of genomic DNA from several macrofungal cultures
[0075] The samples were the mycelial cultures of Lactarius ruficola, Boletus, Pleurotus chinensis and Pleurotus ulmis, numbered 1, 2, 3 and 4, respectively.
[0076] Extraction steps: (1) Put a small amount of mycelium samples into the prepared PCR tubes with ES solution; (2) Put the PCR tubes into the PCR instrument and incubate at 95°C for 10 min; (3 ) After taking it out, add BSA solution 3 times the volume of ES solution, and the extraction is complete; (4) Use spectrophotometer to measure the content of genomic DNA, and the measurement results are as follows: image 3 Shown: Contents are respectively "1" Mycelium culture of Lactaria rufica: 596.58 ng / µL, "2" Mycelia culture of Boletus: 558.38 ng / µL, "3": Mycelia of A. The somatic culture was 494.58 ng / µL, the "4" ulm yellow mycelia culture: 529.58 ng / µL; (5) To verify the DNA extraction effect, the ITS fragment was amplified by PCR, and the a...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More