CN115232810A - Method for extracting total RNA of fishbone - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 37
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 83
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims abstract description 52
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 38
- 239000007788 liquid Substances 0.000 claims abstract description 30
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- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 28
- 239000006228 supernatant Substances 0.000 claims description 26
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 16
- 239000004570 mortar (masonry) Substances 0.000 claims description 16
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- 238000002123 RNA extraction Methods 0.000 claims description 11
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- 238000002474 experimental method Methods 0.000 abstract description 7
- 102000011931 Nucleoproteins Human genes 0.000 abstract description 6
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Abstract
The invention discloses a method for extracting total RNA of fishbone, belonging to the field of biotechnology. The method is suitable for fish bone tissue samples, and on the premise of reducing the activity of RNase in cells and reducing the degradation of total RNA, the nucleoprotein body is completely separated by adding liquid nitrogen, oscillating, ice bath and other modes; washing with ethanol, treating with enzyme-free sterile water and ice bath to obtain high-quality total RNA. The method has the advantages of high extraction efficiency, convenience and rapidness, reduction of the consumption of the RNase removing centrifuge tube and the pipette tip, low price and common reagents, effective reduction of economic cost, reduction of the RNase removing treatment steps of experimental consumables, and great importance for poor experimental environmental conditions or experiment novices.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a method for extracting total RNA of fishbone.
Background
At present, with the rapid development of modern molecular biology technology research, the research on genome RNA has become one of the current research hot directions, but the molecular properties of the genome RNA and the genome DNA are different, the primary structure of the genome RNA is a single-stranded structure, and a double-helix structure is not formed, so that the structural stability of the genome RNA is poor, the degradation phenomenon is easy to occur, and the subsequent experimental research results are directly influenced.
Although there are many kinds of kits for extracting total RNA in the modern market, and the operation described in the specification can achieve a good effect, in the actual operation process of the personnel, many experimental results do not reach the expected effect because of the influence of many factors such as the kind of experimental sample, the experimental operation environment, the experimental operation method, and the laboratory conditions, and especially, the kit is a great obstacle for the experiment novice.
In order to obtain higher-quality total RNA, a large number of repeated experiments are required or a third party is entrusted to extract; in addition, in general, the extraction experiment environment of genome RNA needs to be performed in a strict sterile and enzyme-free environment, the used experiment consumables also need to be subjected to strict enzyme-free treatment, and the cost of the experiment is increased virtually, so that it is very important to find a novel efficient and economic total RNA extraction method.
Disclosure of Invention
The invention aims to provide a method for extracting total RNA from fishbone, which can obtain high-quality total RNA by using sterile environment and only performing two-time circular extraction, and solves the product standard which can be achieved by repeated experiments in the prior art; meanwhile, the technical problem of high requirements on the total RNA extraction experimental environment in the prior art is also solved.
The invention is realized by the following technical scheme:
a method for extracting total RNA of fishbone comprises the following steps:
s1, selecting fresh or frozen fishbone tissues to be crushed within 2-3 min;
s2, placing the crushed fishbone tissue in a mortar on a superclean bench, adding liquid nitrogen, grinding into powder, adding a total RNA extraction reagent, uniformly mixing by oscillation, placing in an ice bath, centrifuging and taking supernatant;
s3, adding chloroform into the supernate, oscillating, carrying out ice bath and centrifuging to obtain a mixture consisting of a colorless water sample layer, an intermediate layer and a red organic chloroform layer from top to bottom;
s4, absorbing the colorless water sample layer substances in the S3, adding isopropanol, mixing uniformly, placing in an ice bath, centrifuging, removing supernatant liquid, then adding ethanol, washing and precipitating, placing in an ice bath, centrifuging, and removing supernatant liquid; washing and precipitating with ethanol for the second time, placing in ice bath, centrifuging, and removing supernatant; standing at room temperature, air drying, adding enzyme-free sterile water for water bath treatment, and immediately performing ice bath treatment to obtain total RNA.
Said invention provides a method for extracting total RNA from fish bone, in said method fresh or frozen sample stored at-80 deg.C is selected, and the fish bone tissue is cut into pieces in short time, then quickly frozen so as to reduce activity of RNase in cell and reduce degradation of total RNA.
In S2, the sterilization process and the operation of the clean bench are performed by the following steps: opening an ultra-clean workbench, disinfecting the ultra-clean workbench surface by using 75% ethanol solution, putting the sterilized mortar, medical forceps, medical scissors, a common centrifuge tube, an RNase-free or common pipette tip, a micro-pipette gun, latex gloves and required reagents into the ultra-clean workbench together, closing a fluorescent lamp, opening an ultraviolet lamp, carrying out ultraviolet irradiation for 15min, and ventilating for 5min.
In the S2, the crushed fishbone tissues are placed in a mortar on a clean bench, liquid nitrogen is added into the process of grinding the fishbone tissues into powder, the grinding time of each tissue sample needs to be strictly controlled within 3min, and the liquid nitrogen is always kept in the mortar in the liquid nitrogen grinding process.
In the above S2, the objective of shaking, mixing and placing in ice bath is to completely separate the nucleoprotein body.
In the above S3, after adding chloroform, the treatment of shaking may be performed by vortexing for 15 seconds or by inverting the mixture for 1.5 min.
In the S4, the purpose of washing and precipitating by using ethanol twice is to remove protein, lipid and other impurity molecules and improve the yield and the purity of RNA.
In the above S4, enzyme-free sterile water is added for water bath treatment in order to sufficiently dissolve RNA; the ice-bath treatment is carried out immediately, the ice-bath time is extremely short, and the preservation environment is required at the same time, so that the degradation of the total RNA obtained by extraction is prevented.
Preferably, the frozen fish bone tissue is preserved at-80 ℃.
Preferably, in the step S2, the time for adding liquid nitrogen for grinding is not more than 3min.
Preferably, in the S2, the dosage ratio of the broken fishbone tissue to the total RNA extraction reagent is 30-60mg:1mL.
Preferably, in the S2, the mixture is uniformly shaken at 2000-2700r/min and is placed for 5-10min under the ice bath condition; centrifuging at 11000-12000r/min for 8-10min at 4-5 deg.C, and collecting supernatant;
the ice bath conditions were 0 ℃.
Preferably, in S3, the dosage ratio of chloroform to total RNA extraction reagent is 0.2-0.22mL:1mL.
The chloroform is used for extracting protein, so that an organic phase and an inorganic phase are rapidly separated, and the layering of the organic phase and the aqueous phase is accelerated.
Preferably, in the S3, the vibration treatment is carried out for 15S-1.5min, the mixture is placed for 2-2.5min under the ice bath condition, and the centrifugation is carried out for 10-15min at 11000-12000r/min under the condition of 4-5 ℃; the ice bath conditions were 0 ℃.
Preferably, in S4, the dosage ratio of the isopropanol to the total RNA extraction reagent is 300-600 μ L:0.5-1mL; standing for 20-30min under ice bath condition; centrifuging at 11000-12000r/min for 10-15min at 4-5 deg.C, and removing supernatant; the ice bath conditions were 0 ℃.
The purpose of using isopropanol was to precipitate RNA as described above.
Preferably, 1.2-1.5mL of 75% ethanol is added into the S4 to wash the precipitate, and the precipitate is placed for 3-5min under the ice bath condition; centrifuging at 11000-12000r/min for 2-3min at 4-5 deg.C, and removing supernatant; the ice bath conditions were 0 ℃.
Preferably, in S4, standing at room temperature for 2-3min, air drying, adding 30-100 μ L of sterile water without enzyme, performing water bath at 55-60 deg.C for 2-3min, immediately performing ice bath for 1-2min to obtain total RNA solution, and storing to-70 deg.C; the ice bath conditions were 0 ℃.
Compared with the prior art, the invention at least has the following technical effects:
the invention provides a method for extracting total RNA from fish bones, which is suitable for fish bone tissue samples, has high extraction efficiency, is convenient and quick, reduces the consumption of an RNase removing centrifuge tube and a pipette tip, has cheap and common reagents, effectively reduces the economic cost, reduces the treatment steps of removing RNase from experimental consumables, and is important for poor experimental environmental conditions or experimental novices.
In the method, samples which are fresh or frozen and stored at-80 ℃ are selected, and the fishbone tissues are required to be cut into pieces in a short time and then are quickly frozen, so that the purpose is to reduce the activity of RNase in cells and reduce the degradation of total RNA. The method comprises the following steps of placing broken fishbone tissues in a mortar on an ultra-clean workbench, adding liquid nitrogen to grind the fishbone tissues into powder, ensuring that the grinding time of each tissue sample is strictly controlled within 3min, and keeping the mortar to have the liquid nitrogen in the liquid nitrogen grinding process. The mixture was shaken and mixed, and then placed in an ice bath to completely separate the nucleoprotein body. The purpose of washing the precipitate with two ethanol is to remove proteins, lipids and other impurity molecules. The yield and purity of RNA are improved. The purpose of adding enzyme-free sterile water for water bath treatment is to fully dissolve RNA; the ice-bath treatment is carried out immediately, the ice-bath time is extremely short, and the preservation environment is required at the same time, so that the degradation of the total RNA obtained by extraction is prevented.
Drawings
FIG. 1 is a schematic process flow diagram of one embodiment of the present invention;
FIG. 2 is a schematic diagram of gel electrophoresis in example 1 of the present invention.
Detailed Description
Embodiments of the present invention will be described in detail with reference to the following examples, but those skilled in the art will understand that the following examples are merely illustrative of the present invention and should not be construed as limiting the scope of the present invention, and that the specific conditions not specified in the examples are carried out according to conventional conditions or conditions suggested by the manufacturer, and that the reagents or equipment used are not specified by the manufacturer, and are all conventional products available through commercial purchase.
The technical scheme of a specific implementation mode of the invention is as follows:
a method for extracting total RNA of fishbone comprises the following steps:
1) Sample pretreatment: collecting fresh or-80 deg.C frozen fishbone tissue in suitable environment, strictly controlling to expose fishbone tissue in external environment within 2-3min, shearing fishbone tissue as much as possible, placing into centrifugal tube, immediately immersing the centrifugal tube containing fishbone tissue sample in liquid nitrogen for cooling and quick freezing. Aiming at reducing the activity of RNase in cells and reducing the degradation of total RNA; the temperature for cryopreservation of the samples should be at-80 ℃.
2) Opening an ultra-clean workbench, sterilizing the ultra-clean workbench surface by using 75% ethanol solution, putting the sterilized mortar, medical forceps, medical scissors, a common centrifuge tube, an RNase-free or common pipette tip, a micro-pipette gun, latex gloves and required reagents into the ultra-clean workbench, closing a fluorescent lamp, opening an ultraviolet lamp, carrying out ultraviolet irradiation for 15min, and ventilating for 5min.
3) 30-60mg of the frozen fishbone tissue is put into a mortar, and liquid nitrogen is added to fully grind the fishbone tissue into tissue powder. Note that: the grinding time of each tissue sample is strictly controlled within 3min, and liquid nitrogen always exists in a mortar during the liquid nitrogen grinding process.
4) Adding 1mL of TRIzolplus into a fishbone tissue sample powder centrifuge tube which is filled with liquid nitrogen and ground into 30-60mg, oscillating at 2000-2700r/min, fully and uniformly mixing, and placing for 5-10min under an ice bath condition to completely separate a nucleoprotein body;
5) Centrifuging at 11000-12000r/min for 8-10min at 4-5 deg.C, and collecting supernatant;
6) 0.2-0.22mL of chloroform was added per 1mL of TRIzolplus. Covering the tube cover tightly, performing vortex oscillation for 15s or reversing the tube cover up and down for 1.5min, standing for 2-2.5min under the ice bath condition, and centrifuging at 11000-12000r/min for 10-15min at 4-5 ℃;
7) The mixture after centrifugation was divided into three layers: the lower layer is a red organic chloroform layer, the middle layer is a colorless water sample layer. Absorbing the upper colorless water sample layer material into a clean centrifugal tube;
8) Adding 600 μ L isopropanol, turning upside down, mixing, and standing under ice bath condition for 20-30min;
9) Centrifuging at 11000-12000r/min for 10-15min at 4-5 deg.C, and removing supernatant;
10 Adding 1.2mL of 75% ethanol by mass percentage to wash the precipitate, turning the precipitate upside down to wash the precipitate, and placing the precipitate for 3min under the ice bath condition;
11 Centrifuging at 11000-12000r/min for 3-5min at 4-5 deg.C, discarding supernatant, and taking care not to lose RNA precipitate;
12 Repeatedly washing the precipitate with 1.2mL of 75% ethanol by mass percent, washing the precipitate in an upside-down manner, placing the precipitate for 3-5min under the ice bath condition, centrifuging the precipitate for 10min at the temperature of 4 ℃ at 12000r/min, and discarding the supernatant;
13 ) standing at room temperature for 2-3min, and air drying. Adding 30-100 μ L of enzyme-free sterile water (RNase free water) in 55-60 deg.C water bath for 2-3min to dissolve RNA, immediately ice-cooling at 0 deg.C for 2min, and instantly separating to obtain RNA solution, and storing at-70 deg.C to prevent degradation.
The selected TRIzolplus is a novel total RNA extraction reagent, and the product is obtained from the following components: the manufacturer GeneNode; the goods number is: 2101B.
Example 1: the extraction process is shown in FIG. 1
A method for extracting total RNA of fishbone comprises the following steps:
1) Sample pretreatment: collecting fresh or-80 deg.C frozen fishbone tissue in suitable environment, strictly controlling to expose fishbone tissue in external environment within 2-3min, shearing fishbone tissue as much as possible, placing into centrifugal tube, immediately immersing the centrifugal tube containing fishbone tissue sample in liquid nitrogen for cooling and quick freezing.
2) Opening an ultra-clean workbench, sterilizing the ultra-clean workbench surface by using 75% ethanol solution, putting the sterilized mortar, medical forceps, medical scissors, a common centrifuge tube, an RNase-free or common pipette tip, a micro-pipette gun, latex gloves and required reagents into the ultra-clean workbench, closing a fluorescent lamp, opening an ultraviolet lamp, carrying out ultraviolet irradiation for 15min, and ventilating for 5min.
3) 30-60mg of the frozen fishbone tissue is put into a mortar, and liquid nitrogen is added to fully grind the fishbone tissue into tissue powder. Note that: the grinding time of each tissue sample is strictly controlled within 3min, and liquid nitrogen always exists in a mortar during the liquid nitrogen grinding process.
4) Adding 1mLTRIZOL plus into a fishbone tissue sample powder centrifuge tube which is filled with liquid nitrogen and ground into 30-60mg, oscillating at 2000-2700r/min and mixing uniformly, and placing for 5min under ice bath condition to completely separate a nucleoprotein body;
5) Centrifuging at 12000r/min for 10min at 4 deg.C, and collecting supernatant;
6) 0.22mL of chloroform was added per 1mL of TRIzolplus. Covering the tube cover tightly, performing vortex oscillation for 15s or turning upside down for 1.5min, standing under ice bath condition for 2.5min, and centrifuging at 12000r/min for 10-15min at 4 deg.C;
7) The mixture after centrifugation was divided into three layers: the lower layer is a red organic chloroform layer, the middle layer is a colorless water sample layer. Absorbing the upper colorless water sample layer material into a clean centrifugal tube;
8) Adding 600 μ L isopropanol, turning upside down, mixing, and standing under ice bath condition for 20-30min;
9) Centrifuging at 12000r/min at 4 deg.C for 10-15min, and discarding the supernatant;
10 Adding 1.2mL of 75% ethanol by mass percentage to wash the precipitate, turning the precipitate upside down to wash the precipitate, and placing the precipitate for 3min under the ice bath condition;
11 ) centrifuging at 12000r/min for 3min at 4 deg.C, discarding the supernatant, taking care not to lose the RNA pellet;
12 Repeatedly washing the precipitate with 1.2mL of 75% ethanol by mass percent, washing the precipitate upside down, placing the precipitate for 3min under the ice bath condition, centrifuging the precipitate for 10min at 4 ℃ at 12000r/min, and removing supernatant;
13 ) standing at room temperature for 3min, and air drying. Adding 30-100 μ L sterile water (RNase free water) with no enzyme at 55-60 deg.C in water bath for 2min, dissolving RNA sufficiently, immediately ice-cooling at 0 deg.C for 2min, instantly separating, and storing the obtained RNA solution to-70 deg.C to prevent degradation.
Example 2: the extraction process is shown in FIG. 1
A method for extracting total RNA of fishbone comprises the following steps:
1) Sample pretreatment: collecting fresh or-80 deg.C frozen fishbone tissue in suitable environment, strictly controlling to expose fishbone tissue in external environment within 2-3min, shearing fishbone tissue as much as possible, placing into centrifugal tube, immediately immersing the centrifugal tube containing fishbone tissue sample in liquid nitrogen for cooling and quick freezing. The temperature for cryopreservation of the samples should be at-80 ℃.
2) Opening an ultra-clean workbench, sterilizing the ultra-clean workbench surface by using 75% ethanol solution, putting the sterilized mortar, medical forceps, medical scissors, a common centrifuge tube, an RNase-free or common pipette tip, a micro-pipette gun, latex gloves and required reagents into the ultra-clean workbench, closing a fluorescent lamp, opening an ultraviolet lamp, carrying out ultraviolet irradiation for 15min, and ventilating for 5min.
3) Taking 30-60mg of the frozen and preserved fishbone tissue into a mortar, adding liquid nitrogen, and fully grinding the fishbone tissue into tissue powder. Note that: the grinding time of each tissue sample is strictly controlled within 3min, and liquid nitrogen always exists in a mortar during the liquid nitrogen grinding process.
4) Adding 1mL of TRIzolplus into a fishbone tissue sample powder centrifuge tube which is filled with liquid nitrogen and ground into 30-60mg, oscillating at 2000-2700r/min, mixing uniformly, and standing for 5-10min under the ice bath condition to completely separate a nucleoprotein body;
5) Centrifuging at 12000r/min for 8min at 4 deg.C, and collecting supernatant;
6) 0.22mL of chloroform was added per 1mL of TRIzolplus. Covering the tube cover tightly, performing vortex oscillation for 15s or turning upside down for 1.5min, standing under ice bath condition for 2.5min, and centrifuging at 12000r/min for 10-15min at 4 deg.C;
7) The mixture after centrifugation was divided into three layers: the lower layer is a red organic chloroform layer, the middle layer is a colorless water sample layer. Absorbing colorless water sample layer substances into a clean centrifugal tube;
8) Adding 600 μ L isopropanol, turning upside down, mixing, and standing under ice bath condition for 20-30min;
9) Centrifuging at 12000r/min for 15min at 4 deg.C, and removing supernatant;
10 Adding 1.2mL of 75% ethanol by mass percentage to wash the precipitate, turning the precipitate upside down to wash the precipitate, and placing the precipitate for 3min under the ice bath condition;
11 ) centrifuging at 12000r/min for 5min at 4 deg.C, discarding the supernatant, and taking care not to lose the RNA precipitate;
12 Repeatedly washing the precipitate with 1.2mL of 75% ethanol by mass percent, washing the precipitate upside down, placing the precipitate for 5min under the ice bath condition, centrifuging the precipitate for 10min at 12000r/min at 4 ℃, and removing supernatant;
13 ) standing at room temperature for 3min, and air drying. Adding 30-100 μ L enzyme-free sterile water (RNase free water) into 55-60 deg.C water bath for 3min, dissolving RNA sufficiently, immediately ice-bathing for 2min, and instantly separating to obtain RNA solution, storing at-70 deg.C, and preventing degradation.
Experimental example:
the gel electrophoresis method verification is carried out by taking the example 1 as an experimental example, and the result shows that:
as shown in FIG. 2, the 1 st and 8 th bands are two 100-5000bp Maker bands, and the remaining 6 bands are extracted RNA bands.
In the figures 2-7, RNA strips 28s,18s and 5s of a first layer, a second layer and a third layer are clear, have no degradation, no protein pollution, better RNA integrity and high extraction quality from top to bottom.
Finally, it should be noted that: the above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A method for extracting total RNA from fishbone is characterized by comprising the following steps:
s1, selecting fresh or frozen fishbone tissues to be crushed within 2-3 min;
s2, placing the crushed fishbone tissue in a mortar on a superclean bench, adding liquid nitrogen, grinding into powder, adding a total RNA extraction reagent, uniformly mixing by oscillation, placing in an ice bath, centrifuging and taking supernatant;
s3, adding chloroform into the supernate, and carrying out oscillation, ice bath and centrifugal treatment to obtain a mixture consisting of a colorless water sample layer, an intermediate layer and a red organic chloroform layer from top to bottom;
s4, absorbing the colorless water sample layer substances in the S3, adding isopropanol, uniformly mixing, placing in an ice bath, centrifuging, removing supernatant liquid, adding ethanol, washing and precipitating, placing in an ice bath, centrifuging, and removing supernatant liquid; washing and precipitating with ethanol for the second time, placing in ice bath, centrifuging, and removing supernatant; standing at room temperature, air drying, adding enzyme-free sterile water for water bath treatment, and immediately performing ice bath treatment to obtain total RNA.
2. The method for extracting total RNA from fish bone according to claim 1, wherein the frozen fish bone tissue is preserved at-80 ℃.
3. The method for extracting total RNA from fish bone according to claim 1, wherein the time for adding liquid nitrogen to grind in S2 is not more than 3min.
4. The method for extracting total RNA from fish bone according to claim 1, wherein in the step S2, the dosage ratio of the crushed fish bone tissue to the total RNA extraction reagent is 30-60mg:1mL.
5. The method for extracting total RNA from fishbone according to claim 1, wherein in S2, the fishbone is uniformly mixed by shaking at 2000-2700r/min and placed for 5-10min under ice bath condition; centrifuging at 11000-12000r/min for 8-10min at 4-5 deg.C, and collecting supernatant;
the ice bath conditions were 0 ℃.
6. The method for extracting total RNA from fish bones as claimed in claim 1, wherein in S3, the dosage ratio of chloroform to total RNA extraction reagent is 0.2-0.22mL:1mL.
7. The method for extracting total RNA from fish bone according to claim 1, wherein in S3, the fish bone is treated by shaking for 15S-1.5min, placed for 2-2.5min under ice bath condition, and centrifuged for 10-15min at 11000-12000r/min at 4-5 ℃;
the ice bath conditions were 0 ℃.
8. The method for extracting total RNA from fish bone according to claim 1, wherein in the S4, the dosage ratio of isopropanol to the total RNA extraction reagent is 300-600 μ L:0.5-1mL;
standing for 20-30min under ice bath condition; centrifuging at 11000-12000r/min for 10-15min at 4-5 deg.C, and removing supernatant;
the ice bath conditions were 0 ℃.
9. The method for extracting total RNA from fish bones as claimed in claim 1, wherein 1.2-1.5mL of 75% ethanol is added to S4 to wash the precipitate, and the precipitate is placed for 3-5min under ice bath conditions; centrifuging at 11000-12000r/min for 2-3min at 4-5 deg.C, and removing supernatant;
the ice bath conditions were 0 ℃.
10. The method for extracting total RNA from fish bones as claimed in claim 1, wherein in S4, the fish bones are placed at room temperature for 2-3min, dried, added with 30-100 μ L of sterile water without enzyme, bathed in water at 55-60 ℃ for 2-3min, immediately bathed in ice for 1-2min to obtain total RNA solution, and stored to-70 ℃;
the ice bath conditions were 0 ℃.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103642797A (en) * | 2013-12-11 | 2014-03-19 | 华中农业大学 | Extraction method of total RNA of intermuscular bone of megalobrama amblycephala |
| CN107904233A (en) * | 2017-12-31 | 2018-04-13 | 贵州大学 | A kind of method for total RNA from animal tissues extraction |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103642797A (en) * | 2013-12-11 | 2014-03-19 | 华中农业大学 | Extraction method of total RNA of intermuscular bone of megalobrama amblycephala |
| CN107904233A (en) * | 2017-12-31 | 2018-04-13 | 贵州大学 | A kind of method for total RNA from animal tissues extraction |
Non-Patent Citations (1)
| Title |
|---|
| 郑纺等: ""改良TRIZOL 法从矿化骨组织中提取总RNA"", 《天津医药》, 31 December 2007 (2007-12-31), pages 753 - 754 * |
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