Extraction method of total rna from the adults of Leafhopper chrysalis
A technology of small green leafhopper and extraction method, which is applied in the direction of DNA preparation and recombinant DNA technology, and can solve the problems of difficult extraction of total RNA and insufficient purity.
Inactive Publication Date: 2016-06-15
信阳市农业科学院
View PDF5 Cites 0 Cited by
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
[0008] The purpose of the present invention is to address the deficiencies in the prior art, thereby providing the extraction method of the total RNA of the green leafhopper adult, to solve the total RNA extraction of such small insects as the green leafhopper adult in the prior art Difficulty and the problem that the purity cannot meet the requirements
Method used
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View moreImage
Smart Image Click on the blue labels to locate them in the text.
Smart ImageViewing Examples
Examples
Experimental program
Comparison scheme
Effect test
Embodiment Construction
[0029] The technical solutions of the present invention will be described in further detail below through specific implementation methods.
[0030] The method for extracting the total RNA of the green leafhopper adult larvae comprises the following steps:
[0031] Step 1. Collect the adults of the green leafhopper in a 1.5ml centrifuge tube, and heat-shock the adults of the green leafhopper at a temperature of 34-36°C for 50-70 minutes, so that the adults of the green leafhopper are heat-shocked and comatose , and then immediately place the centrifuge tube in liquid nitrogen;
[0032] Step 2. Take out the centrifuge tube from the liquid nitrogen and immediately add the total RNA extraction solution RNAisoPlus, fully homogenize with a plastic grinding rod, and it becomes transparent without particles, then add the total RNA extraction solution RNAisoPlus to a sufficient amount, and then use a 1ml pipette Beat the slurry several times;
[0033] Step 3. Place the homogenate sam...
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More PUM
Login to View More Abstract
The invention provides a method for extracting total ribonucleic acid (RNA) of an empoasca onukii matsuda imago. The method comprises the following steps: 1, performing heat shock treatment on the empoasca onukii matsuda imago, and putting the empoasca onukii matsuda imago into liquid nitrogen; 2, adding a total RNA extraction liquid RNAisoPlus into a centrifuge tube, and homogenating; 3, placing a homogenated sample at the room temperature, and centrifuging; 4, absorbing the supernate, and transferring the supernate into another centrifuge tube; 5, adding chloroform into the supernate, standing at the room temperature, and centrifuging; 6, taking out the centrifuge tube, sucking the supernate, transferring the supernate into a third centrifuge tube, adding isopropanol, standing, and centrifuging; 7, carefully abandoning the supernate, adding ethanol, washing, subsequently centrifuging, abandoning the ethanol, drying, and adding a proper amount of RNase-free water to obtain the total RNA with high purity. The method provided by the invention is simple and effective, and the problems that the total RNA is easy to degrade in the extraction process and the total RNA with high purity is not easy to extract are solved.
Description
technical field [0001] The invention relates to a method for extracting total RNA of small leafhopper insects, in particular to a method for extracting total RNA of adult green leafhoppers. Background technique [0002] The small green leafhopper Empoascaonukii Matsuda is a major pest of tea trees worldwide, and it is also the most widely distributed and most serious tea pest in China. In recent years, with the continuous increase of the planting area of tea trees, the insect has continued to spread, seriously affecting the yield and quality of tea. The buds and leaves damaged by this insect are fragile during the processing process, resulting in a burnt smell, the finished tea leaves are loose, and the broken pieces increase. After brewing, the soup color is cloudy, the taste is bitter, and the aroma is abnormal. In general, summer and autumn tea production will be reduced by 10%-15% in the year of damage, and the production reduction will be more than 50% in the year of...
Claims
the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More Application Information
Patent Timeline
Login to View More Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 乔利卢兆成刘祥臣吕立哲潘兹亮丰大清李慧龙扶定李平王强
Owner 信阳市农业科学院