CN112725335A - Application of filter material in RNA extraction and method for extracting RNA by using filter material - Google Patents
Application of filter material in RNA extraction and method for extracting RNA by using filter material Download PDFInfo
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Abstract
The invention relates to the technical field of biology, in particular to application of a filter material in RNA extraction and a method for extracting RNA by using the filter material. The filtering material is a gauze bag or gauze with the aperture of 1-3 mm. According to the RNA extraction method of the filtering material, the RNA of the plant with high cellulose content can be effectively extracted, and the purity and concentration of the extracted RNA are improved.
Description
Technical Field
The invention relates to the technical field of biology, in particular to application of a filter material in RNA extraction and a method for extracting RNA by using the filter material.
Background
DNA, RNA and proteins are three important biological macromolecules that are the molecular basis for life phenomena. Genetic information of DNA determines major traits of life, and mRNA plays an important role in information transmission. The other two major classes of RNA, rRNA and tRNA, also play irreplaceable important roles in protein biosynthesis.
When extracting RNA from different plants, it is difficult to extract RNA from different plants using the same method because different parts of different plants or the same plant have their own characteristics. In the case of plants or plant parts with a high cellulose content, the prior art does not satisfy the purity and concentration requirements for RNA extraction.
Disclosure of Invention
The invention aims to provide application of a filter material in RNA extraction and a method for extracting RNA by using the filter material.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides application of a filter material in RNA extraction, wherein the filter material is a gauze bag or gauze.
Preferably, the aperture of the mesh of the gauze bag or the gauze is 1-3 mm.
Preferably, the RNA is RNA in plant shoots.
Preferably, the RNA is RNA of plant roots.
The invention also provides a method for extracting RNA by using the filter material, which comprises the following steps:
(1) grinding plant tissue in a filter material to obtain powder containing RNA;
(2) adding lysis solution into the powder containing RNA, centrifuging, and taking supernatant to obtain solution containing RNA;
(3) the RNA-containing solution is purified to obtain RNA.
Preferably, the grinding is carried out under liquid nitrogen;
the mass volume ratio of the plant tissue to the lysis solution is 1-2 mg:10 mu L;
the lysis solution is buffer (SL) and mercaptoethanol;
the volume ratio of SL to mercaptoethanol is 18-20: 1.
Preferably, the centrifugal rotation speed in the step (2) is 12000-13400 rpm; the centrifugation time is 1-3 min.
The invention provides application of a filter material in RNA extraction and a method for extracting RNA by using the filter material. When the filter material is used for RNA extraction, the interference of plant RNA fibrous tissues can be reduced, plant materials are prevented from splashing when liquid nitrogen is ground, and the purity and concentration of RNA can be effectively improved.
Detailed Description
The invention provides application of a filter material in RNA extraction, wherein the filter material is a gauze bag or gauze.
In the invention, the aperture of the mesh of the gauze bag or the gauze is 1-3 mm, preferably 2 mm.
In the present invention, the RNA is RNA in plant shoots.
In the present invention, the RNA is RNA of plant roots.
In the present invention, the rosa plant is a plant having a high cellulose content.
The invention also provides a method for extracting RNA by using the filter material, which comprises the following steps:
(1) grinding plant tissue in a filter material to obtain powder containing RNA;
(2) adding lysis solution into the powder containing RNA, centrifuging, and taking supernatant to obtain solution containing RNA;
(3) the RNA-containing solution is purified to obtain RNA.
In the invention, the filtering material is placed on a grinder vessel for grinding during grinding.
In the present invention, the grinding vessel is preferably a mortar.
In the present invention, the milling is milling performed under liquid nitrogen.
In the present invention, the powder of the RNA-containing powder is a powder filtered out by a filter material.
In the invention, the mass-volume ratio of the plant tissue to the lysis solution is 1-2 mg:10 muL, preferably 1.5mg:10 muL;
the lysis solution is buffer (SL) and mercaptoethanol;
the SL consists of 8-hydroxyquinoline, guanidinium isothiocyanate and beta-mercaptoethanol;
the volume ratio of SL to mercaptoethanol is 18-20: 1, and is preferably 19: 1.
In the invention, the centrifugal rotation speed in the step (2) is 12000-13400 rpm; preferably 12700 rpm; the centrifugation time is 1-3 min, preferably 2 min.
In the present invention, the purification in the step (3) comprises the steps of:
(3.1) transferring the solution containing RNA to an RNase-FreeSpin Column CS Set (CS) filter Column, centrifuging at 12000-13400 rpm for 1-3 min, and taking the supernatant to obtain an intermediate 1;
(3.2) adding absolute ethyl alcohol with the volume 0.3-0.5 time of that of the intermediate 1 into the intermediate 1, uniformly mixing, transferring the mixture into an adsorption Column RNase-Freespin Column CR3 set (CR3) with a collecting pipe, centrifuging at 12000-13400 rpm for 13-17 s, and pouring waste liquid in the collecting pipe to obtain an intermediate 2;
(3.3) adding sodium acetate into an adsorption column CR3 containing the intermediate 2, centrifuging at 12000-13400 rpm for 13-17 s, and pouring off waste liquid in a collecting pipe to obtain an intermediate 3; the mass-volume ratio of the plant tissues to the sodium acetate is 1-2 mg: 6-8 mu L;
(3.4) adding a protease inhibitor cocktail into an adsorption column CR3 containing the intermediate 3, standing at the temperature of 25-28 ℃ for 13-17 min to obtain an intermediate 4; the mass-volume ratio of the plant tissue to the protease inhibitor cocktail is 1-2 mg: 1.2-2 mu L;
(3.5) adding sodium acetate into the adsorption column CR3 containing the intermediate 4 again, centrifuging at 12000-13400 rpm for 13-17 s, and pouring off waste liquid in the collection tube to obtain an intermediate 5; the mass-volume ratio of the plant tissues to the sodium acetate is 1-2 mg: 6-8 mu L;
(3.6) adding an absolute ethyl alcohol rinsing liquid into an adsorption column CR3 containing the intermediate 5, centrifuging at 12000-13400 rpm for 13-17 s, pouring off waste liquid in a collecting pipe, repeating the step for 1-3 times, adding an absolute ethyl alcohol solution into the last time, and centrifuging at 12000-13400 rpm for 1-3 min to obtain an intermediate 6; the mass-volume ratio of the plant tissue to the absolute ethyl alcohol is 1-2 mg: 8-12 mu L;
(3.7) placing the adsorption column CR3 containing intermediate 6 into a container, and adding RNase-Free ddH dropwise into the middle of the adsorption membrane of adsorption column CR32O, standing at 25-28 ℃ for 1-3 min at 12000-13400 rpm, and centrifuging for 1-3 min to obtain RNA; the plant tissue and RNase-Free ddH2The mass-to-volume ratio of O is 5-10 mg: 3-5 μ L.
In the invention, the centrifugal rotation speed of the steps (3.1), (3.2), (3.3), (3.5), (3.6) and (3.7) is 12000-13400 rpm, and preferably 12700 rpm.
In the invention, the centrifugation time of the steps (3.1), (3.6) and (3.7) is 1-3 min, and preferably 2 min.
In the invention, absolute ethyl alcohol with the volume of 0.3-0.5 time of that of the intermediate 1 is added in the step (3.2), and the absolute ethyl alcohol with the volume of 0.4 time is preferably added.
In the present invention, the centrifugation in the steps (3.2), (3.3), (3.5) and (3.6) is carried out for 13 to 17 seconds, preferably for 14 to 16 seconds, and more preferably for 15 seconds.
In the invention, the mass-to-volume ratio of the plant tissue and the sodium acetate in the step (3.3) is 1-2 mg: 6-8 mu L, preferably 1.5mg: 7 μ L.
In the invention, the temperature of the step (3.4) is 25-28 ℃, preferably 26-27 ℃, and more preferably 26.5 ℃.
In the invention, the step (3.4) is left for 13-17 min, preferably 14-16 min, and more preferably 15 min.
In the invention, the mass-to-volume ratio of the plant tissue and the protease inhibitor cocktail in the step (3.4) is 1-2 mg: 1.2-2 muL, preferably 1.5mg: 1.6. mu.L.
In the invention, the mass-to-volume ratio of the plant tissue and the sodium acetate in the step (3.5) is 1-2 mg: 6-8 mu L, preferably 1.5mg: 7 μ L.
In the present invention, the step (3.6) is repeated 1 to 3 times, preferably 2 times.
In the invention, the mass-to-volume ratio of the plant tissue and the absolute ethyl alcohol in the step (3.6) is 1-2 mg: 8-12 mu L, preferably 1.5mg:10 μ L.
In the invention, the temperature of the step (3.7) is 25-28 ℃, preferably 26-27 ℃, and more preferably 26.5 ℃.
In the invention, the step (3.7) is left for 1-3 min, preferably 2 min.
In the present invention, the step (3.7) of the tissue of the plant is incubated with RNase-Free ddH2The mass-to-volume ratio of O is 5-10 mg: 3-5 μ L, preferably 7.5 mg: 4 μ L.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Weighing 1g of plum blossom tender branches, and putting the plum blossom tender branches into a mortar sleeved with a gauze bag, wherein the diameter of meshes of the gauze bag is 3 mm. Grinding the branches into powder under the condition of liquid nitrogen, and removing the gauze bag. Adding 7000 mu L of lysate prepared from SL and mercaptoethanol in a volume ratio of 19:1 into the powder, immediately mixing the lysate with vortex oscillation at 13000rpm, and centrifuging for 3 min; sucking supernatant, transferring to a CS filter column with a collecting pipe, performing centrifugation at 13000rpm for 2 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.5 time that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column at 13000rpm, centrifuging for 17s, and pouring off waste liquid in the collection pipe; adding 5000 μ L sodium acetate into adsorption column CR3 at 13000rpm, centrifuging for 17s, and removing waste liquid in the collection tube; adding 1200 μ L protease inhibitor cocktail into adsorption column CR3, and standing at 25 deg.C for 15 min; adding 6000 mu L of sodium acetate into the adsorption column CR3, centrifuging at 13000rpm for 17s, and pouring off waste liquid in the collection tube; adding 8000 mul of absolute ethyl alcohol rinsing liquid into an adsorption column CR3, centrifuging at 13000rpm for 17s, pouring the waste liquid in the collecting pipe, repeating the step for 2 times, centrifuging at 13000rpm for 2min for the last time, and pouring the waste liquid in the collecting pipe; suspending and dripping 400 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, left at 25 ℃ for 2min, and centrifuged at 13000rpm for 1min to obtain the RNA solution of example 1.
Determination of OD in RNA solution in example 1230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated. The results are shown in Table 1.
Example 2
100mg of rose old branches are weighed and put into a mortar with gauze, and the diameter of gauze meshes is 2 mm. Grinding rose old branches into powder under the condition of liquid nitrogen, and removing gauze. Adding SL and mercaptoethanol into the powder in a volume ratio of 20:1, immediately carrying out vortex oscillation and uniform mixing on 850 mu L of lysate prepared by the method, carrying out centrifugation at 11000rpm for 3 min; sucking supernatant, transferring to a CS filter column with a collecting pipe, centrifuging at 11000rpm for 3 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.3 time of that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column at 11000rpm, centrifuging for 13s, and pouring off waste liquid in the collection pipe; adding 800 μ L sodium acetate into the adsorption column CR3 at 11000rpm, centrifuging for 13s, and pouring off the waste liquid in the collection tube; adding 200 μ L protease inhibitor cocktail into adsorption column CR3, standing at 28 deg.C for 13 min; adding 600 mu L of sodium acetate into an adsorption column CR3, centrifuging at 11000rpm for 13s, and pouring waste liquid in a collecting pipe; adding 1000 μ L of anhydrous ethanol rinsing solution into adsorption column CR3, centrifuging at 11000rpm for 13s, pouring off waste liquid in the collection tube, repeating the step for 1 time, centrifuging at 11000rpm for 3min for the last time, and pouring off waste liquid in the collection tube; suspending and dripping 40 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, standing at 28 ℃ for 3min, and centrifuging at 11000rpm for 2min to obtain the RNA solution of example 2.
Determination of OD in RNA solution in example 2230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated. The results are shown in Table 1.
Example 3
Weighing 1g of orchid roots, and putting the orchid roots into a mortar sleeved with a gauze bag, wherein the diameter of meshes of the gauze bag is 2 mm. Grinding the orchid root into powder under the condition of liquid nitrogen, and removing the gauze bag. Adding 10000 microliter of lysate prepared from SL and mercaptoethanol in a volume ratio of 18:1 into the powder, immediately and uniformly mixing by vortex oscillation at 12500rpm, and centrifuging for 2 min; sucking supernatant, transferring to a CS filter column with a collecting pipe, and centrifuging at 12500rpm for 2 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.4 time that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column at 12500rpm, centrifuging for 13s, and pouring out waste liquid in the collection pipe; adding 6000 μ L sodium acetate into the adsorption column CR3 at 12500rpm, centrifuging for 17s, and removing waste liquid in the collection tube; suction nozzleAdding 1500 μ L protease inhibitor cocktail into attached column CR3, standing at 28 deg.C for 13 min; adding 8000 mu L sodium acetate into an adsorption column CR3, 12500rpm, centrifuging for 15s, and pouring off waste liquid in a collecting pipe; adding 10000 μ L of anhydrous ethanol rinsing solution into an adsorption column CR3, centrifuging at 12500rpm for 13s, pouring off waste liquid in a collecting pipe, repeating the step for 3 times, centrifuging at 12500rpm for 1min for the last time, and pouring off waste liquid in the collecting pipe; suspending and dripping 400 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, left at 25 ℃ for 1min and centrifuged at 12500rpm for 3min to obtain the RNA solution of example 3.
Determination of OD in RNA solution in example 3230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated. The results are shown in Table 1.
Example 4
Weighing 1g of old pepper branches, and putting the old pepper branches into a mortar sleeved with a gauze bag, wherein the diameter of meshes of the gauze bag is 3 mm. Grinding the old branches of the hot peppers into powder under the condition of liquid nitrogen, and removing the gauze bag. Adding 6000 μ L of lysate prepared from SL and mercaptoethanol at a volume ratio of 18:1 into the powder, immediately mixing the lysate with the powder by vortex oscillation at 13400rpm, and centrifuging for 1 min; sucking supernatant, transferring to a CS filter column with a collecting pipe, rotating at 13400rpm, and centrifuging for 2 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.5 time that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column, carrying out 13400rpm centrifugation for 14s, and pouring off waste liquid in the collection pipe; adding 7500 μ L sodium acetate into adsorption column CR3, rotating at 13400rpm, centrifuging for 14s, and pouring off waste liquid in the collecting tube; adding 1800 μ L protease inhibitor cocktail into adsorption column CR3, and standing at 26 deg.C for 15 min; adding 7500 μ L sodium acetate into adsorption column CR3, rotating at 13400rpm, centrifuging for 14s, and pouring off waste liquid in the collecting tube; adding 8500 mu L of absolute ethyl alcohol rinsing liquid into an adsorption column CR3, centrifuging at 13400rpm for 14s, pouring waste liquid in a collecting pipe, repeating the step for 1 time, centrifuging at 13400rpm for 2min for the last time, and pouring the waste liquid in the collecting pipe; suspending and dripping 400 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, left at 25 ℃ for 1min, and centrifuged at 13400rpm for 1min to obtain the RNA solution of example 4.
Determination of OD in RNA solution in example 4230、OD260And OD280The numerical value of (c). Calculating the sum of the concentrations of RNAAnd (4) content. The results are shown in Table 1.
Example 5
Weighing 1g of old pine branches, and putting the old pine branches into a mortar sleeved with a gauze bag, wherein the diameter of the meshes of the gauze bag is 1 mm. Grinding the old branches of the hot peppers into powder under the condition of liquid nitrogen, and removing the gauze bag. Adding 8000 μ L of lysate prepared from SL and mercaptoethanol at a volume ratio of 20:1 into the powder, immediately mixing by vortex oscillation at 12700rpm, and centrifuging for 3 min; sucking supernatant, transferring to CS filter column with collecting tube, centrifuging at 12700rpm for 3 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.3 time that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column, centrifuging at 12700rpm for 17s, and pouring off waste liquid in the collection pipe; adding 6800 μ L sodium acetate into adsorption column CR3, centrifuging at 12700rpm for 17s, and removing waste liquid in the collection tube; adding 1600 μ L protease inhibitor cocktail into adsorption column CR3, and standing at 27 deg.C for 15 min; adding 6800 μ L sodium acetate into adsorption column CR3, centrifuging at 12700rpm for 17s, and removing waste liquid in the collection tube; adding 9000 μ L of anhydrous ethanol rinsing solution into adsorption column CR3, centrifuging at 12700rpm for 17s, removing waste liquid in the collection tube, repeating the step for 2 times, centrifuging at 12700rpm for 2min for the last time, and removing waste liquid in the collection tube; suspending and dripping 400 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, standing at 26 ℃ for 3min, and centrifuging at 12700rpm for 2min to obtain the RNA solution of example 5.
Determination of OD in RNA solution in example 5230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated. The results are shown in Table 1.
Example 6
Weighing 1g of old peony branches, and putting the old peony branches into a mortar sleeved with a gauze bag, wherein the diameter of meshes of the gauze bag is 3 mm. Grinding old peony branches into powder under the condition of liquid nitrogen, and removing the gauze bag. Adding 7500 μ L of lysate prepared from SL and mercaptoethanol at a volume ratio of 19:1 into the powder, immediately mixing by vortex oscillation at 12700rpm, and centrifuging for 3 min; sucking supernatant, transferring to CS filter column with collecting tube, centrifuging at 12700rpm for 3 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.4 time that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column, centrifuging at 12700rpm for 16s, and pouring off waste liquid in the collection pipe; into adsorption column CR3Adding 7300 μ L sodium acetate, centrifuging at 12700rpm for 16s, and removing waste liquid in the collecting tube; adding 2000 μ L protease inhibitor cocktail into adsorption column CR3, and standing at 26 deg.C for 13 min; adding 7300 μ L sodium acetate into adsorption column CR3, centrifuging at 12700rpm for 16s, and removing waste liquid in the collection tube; adding 12000 μ L of anhydrous ethanol rinsing solution into adsorption column CR3, centrifuging at 12700rpm for 16s, pouring out waste liquid in the collection tube, repeating the step for 2 times, centrifuging at 12700rpm for 3min for the last time, and pouring out waste liquid in the collection tube; suspending and dripping 400 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, left at 27 ℃ for 2min and centrifuged at 12700rpm for 1min to obtain the RNA solution of example 6.
Determination of OD in RNA solution in example 6230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated. The results are shown in Table 1.
Example 7
Weighing 1g of rose tender branches, and putting the rose tender branches into a mortar sleeved with a gauze bag, wherein the diameter of the meshes of the gauze bag is 2 mm. Grinding rose twigs into powder under the condition of liquid nitrogen, and removing the gauze bag. Adding 9000 μ L of lysate prepared from SL and mercaptoethanol at a volume ratio of 19:1 into the powder, immediately mixing by vortex oscillation at 13000rpm, and centrifuging for 2 min; sucking supernatant, transferring to a CS filter column with a collecting pipe, performing centrifugation at 13000rpm for 2 min; sucking the supernatant, slowly adding absolute ethyl alcohol with the volume of 0.3 time that of the supernatant, uniformly mixing, transferring into a CR3 adsorption column at 13000rpm, centrifuging for 16s, and pouring off waste liquid in the collection pipe; 6500 μ L sodium acetate is added into the adsorption column CR3 at 13000rpm, centrifuged for 16s, and waste liquid in the collection tube is poured off; adding 1700 μ L protease inhibitor cocktail into adsorption column CR3, and standing at 25 deg.C for 17 min; 6500 μ L sodium acetate is added into the adsorption column CR3 at 13000rpm, centrifuged for 16s, and waste liquid in the collection tube is poured off; adding 10000 μ L of absolute ethyl alcohol rinsing liquid into an adsorption column CR3, centrifuging at 13000rpm for 16s, pouring off waste liquid in a collecting pipe, repeating the step for 1 time, centrifuging at 13000rpm for 2min for the last time, and pouring off the waste liquid in the collecting pipe; suspending and dripping 400 mu LRNase-Free ddH to the middle position of an adsorption film of the adsorption column2O, left at 25 ℃ for 2min, and centrifuged at 13000rpm for 2min to obtain the RNA solution of example 7.
Assay example 7 in RNA solutionOD230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated. The results are shown in Table 1.
Comparative examples 1 to 7
The RNA in the plum blossom young and tender branches, rose old branches, orchid roots, pepper old branches, pine old branches, peony old branches and rose tender branches are extracted according to the same method as in the embodiments 1 to 7, and only the gauze bags in the embodiments 1 to 7 are not adopted in the comparative examples 1 to 7, which is different from the steps in the embodiments 1 to 7. After RNA extraction, OD in solutions of comparative examples 1 to 7 was measured230、OD260And OD280The numerical value of (c). The concentration and amount of RNA were calculated to compare the effect of the filter material in RNA extraction. The results are shown in Table 1.
TABLE 1 purity, concentration and content of RNA for each treatment
| Method | OD260/OD280 | OD260/OD230 | Concentration (ng/. mu.L) | Total amount (μ g) |
| Example 1 | 1.91 | 2.11 | 744.3 | 37.215 |
| Example 2 | 1.87 | 2.08 | 225.6 | 11.28 |
| Example 3 | 1.92 | 7.80 | 17.6 | 88 |
| Example 4 | 1.91 | 2.32 | 424.3 | 212.15 |
| Example 5 | 1.8 | 2.56 | 1025.5 | 512.75 |
| Example 6 | 1.92 | 2.04 | 648.5 | 324.25 |
| Example 7 | 2.01 | 2.62 | 225.6 | 112.8 |
| Comparative example 1 | 1.21 | 0.64 | 89.3 | 4.465 |
| Comparative example 2 | 1.57 | 0.11 | 23.9 | 1.195 |
| Comparative example 3 | 2.09 | 1.29 | 78.4 | 39.2 |
| Comparative example 4 | 2.10 | 2.36 | 330.6 | 165.3 |
| Comparative example 5 | 1.68 | 2.72 | 620.5 | 310.25 |
| Comparative example 6 | 0.96 | 1.58 | 306.5 | 153.25 |
| Comparative example 7 | 2.04 | 2.54 | 224.1 | 112.05 |
The results in Table 1 show the OD of RNA extracted in examples 1 to 7260/OD280All are between 1.8 and 2.0, OD260/OD230All are greater than 2.0, which indicates that the RNAs extracted in examples 1-7 have neither protein nor carbohydrate contamination, while the RNAs extracted in comparative examples 1-7 have OD260/OD280Less than 1.8 or greater than 2.0, indicating protein contamination. As can be seen from the results of RNA concentrations and contents shown in Table 1, the concentrations and contents of the RNAs obtained in examples 1 to 7 were higher than those of the corresponding RNAs extracted without using the gauze bag.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (7)
1. The application of the filter material in the extraction of RNA is characterized in that the filter material is a gauze bag or gauze.
2. The use of claim 1, wherein the mesh bag or gauze has a mesh size of 1 to 3 mm.
3. Use according to claim 1 or 2, wherein the RNA is RNA in plant shoots.
4. Use according to claim 1 or 2, wherein said RNA is RNA of plant roots.
5. A method for extracting RNA by using a filter material is characterized by comprising the following steps:
(1) grinding plant tissue in a filter material to obtain powder containing RNA;
(2) adding lysis solution into the powder containing RNA, centrifuging, and taking supernatant to obtain solution containing RNA;
(3) the RNA-containing solution is purified to obtain RNA.
6. The method according to claim 5, characterized in that the grinding is carried out under liquid nitrogen conditions;
the mass volume ratio of the plant tissue to the lysis solution is 1-2 mg:10 mu L;
the lysis solution is buffer (SL) and mercaptoethanol;
the volume ratio of SL to mercaptoethanol is 18-20: 1.
7. The method according to claim 6, wherein the centrifugal rotation speed in the step (2) is 12000-13400 rpm; the centrifugation time is 1-3 min.
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