CN108611347A - A kind of extracting method of the black yellow striped skin RNA of middle Warsaw loach - Google Patents
A kind of extracting method of the black yellow striped skin RNA of middle Warsaw loach Download PDFInfo
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- CN108611347A CN108611347A CN201810432740.0A CN201810432740A CN108611347A CN 108611347 A CN108611347 A CN 108611347A CN 201810432740 A CN201810432740 A CN 201810432740A CN 108611347 A CN108611347 A CN 108611347A
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Abstract
The invention discloses the extracting methods of the black yellow striped skin RNA of middle Warsaw loach a kind of, include the following steps：Equipment pre-processes, peeling processing, extraction step, quality testing.This method takes short, and easy to operate, at low cost, stability is good, can solve the problems such as previous fish skin tissue RNA extractions are difficult, and quality is low, low output.
Description
Technical field
The invention belongs to gene engineering technology fields, specifically, being related to a kind of black yellow striped skin RNA's of middle Warsaw loach
Extracting method.
Background technology
Middle Warsaw loach is the distinctive rare fish of China, eats and watches, and has prodigious development prospect.As ornamental
The research of aspect, centering Warsaw loach striped is also seldom, and one of the main reasons is that the skin histology sample acquisition of fish is inconvenient, and RNA's carries
Take difficulty.Since its skin is connect with musculature, toughness foot grind away is difficult in addition, seriously affects and extracts height on therefrom Warsaw loach
Quality skin RNA.Routine sampling grind away method is not fully applicable in middle Warsaw loach skin RNA extractions.Especially to divide
In the experiment for indescribably taking black streaking skin and yellow striped skin, routine sampling method easily causes skin RNA drops due to taking
Solution and the insufficient unsuccessful situation of extraction of grind away.
Invention content
In view of this, the present invention is directed to above-mentioned problem, the extraction of the black yellow striped skin RNA of middle Warsaw loach a kind of is provided
Method, method of the invention can be answered with the black yellow striped skin high quality RNA of Warsaw loach in successfully being extracted in the short time and successfully
It is sequenced for high-throughput transcript profile.
In order to solve the above-mentioned technical problem, the invention discloses the extraction sides of the black yellow striped skin RNA of middle Warsaw loach a kind of
Method includes the following steps：Equipment pre-processes, peeling processing, extraction step, quality testing.
Further, the equipment in step 1, which pre-processes, is specially：The dissecting tool that will be used before extraction as solution plane cut,
Scalpel, tweezers etc. and spoon, mortar are put into high-pressure sterilizing pot sterilizing after being wrapped up with masking foil, and are dried and are pre-chilled.
Further, the peeling in step 2, which is handled, is specially：
Step 2.1, operating personnel need band mask and disposable powder-free gloves, are sprayed with alcohol watering can and gloves and wipe dry, made
Replace new gloves in due course in；
Step 2.2, fish-skin stripping：Since middle Warsaw loach is alepidote, take skin without removing scale；First use 40mg/L MS-
222 solution carry out fish to anaesthetize and break neck, then cut pectoral fin with dissecting scissors, facilitate skinning operations；Fish maw, edge are broken along ventrimeson
Fish head is cut at gill cover edge, then separates fish-skin from neck with scalpel, clamps fish-skin with tweezers, cooperation scalpel slowly will be complete
Whole fish-skin strips out, and rejects a small amount of musculature adhered on fish-skin, and complete fish-skin is laid on masking foil rapidly, liquid is used in combination
Nitrogen freezes；
Step 2.3, striped sample collection：Due to fish-skin softness flexible, it is desirable to it is directly separated black yellow striped and is not easy to,
So freezed on masking foil fish-skin using liquid nitrogen, then the texture with scalpel along black yellow striped is directly cut；
It is operated in the case of liquid nitrogen frozen, can quickly separate the black yellow striped of fish-skin, also can guarantee that its RNA is non-degradable.Then will
Striped skin is packed into 2ml cryopreservation tubes, is put into liquid nitrogen.
Further, the extraction step in step 3 is specially：Middle Warsaw loach striped skin is pre-chilled and complete equipped with liquid
It is ground rapidly in the mortar of nitrogen, side is ground at this time, while being shredded skin histology with dissecting scissors, more the thinner fritter the better, is further continued for
Grinding is until it is in powdered；It takes 60-80mg skin histology powder to be added with the spoon of Liquid nitrogen precooler and has filled the Trizol of 1ml
In the EP pipes of liquid, fully oscillation is uniformly mixed, and is put into 12000g, and supernatant is transferred to new centrifuge tube by 4 DEG C of centrifugation 5min；
1/5 volume of chloroform CHCl is added in previous step centrifuge tube3, 15s vortex mixings, after standing 5min, 12000g, 4 DEG C of centrifugations
15min；Supernatant is transferred to new centrifuge tube；It is primary to repeat previous step, preceding once to act as extracting, latter step is to wash
It is de-, improve RNA purity；It is transferred to new centrifuge tube, isometric isopropanol is added, mixing is stored at room temperature 10min, 12000g, 4 DEG C,
Centrifuge 10min；Supernatant is abandoned, 1ml ice ethyl alcohol is added in precipitation, 12000g, centrifuges 5min by 4 DEG C；Supernatant is abandoned, it is drying precipitated,
It is dissolved in appropriate DEPC water.
Compared with prior art, the present invention can be obtained including following technique effect：
1) the present invention is based on sample pre-processings and RNA extracts reagents in a disguised form to utilize, and develops a kind of for fish skin
The extracting method of RNA.This method takes short, and easy to operate, at low cost, stability is good, can solve previous fish skin tissue RNA
The problems such as extraction is difficult, and quality is low, low output.
2) the middle Warsaw loach skin RNA yield of this method extraction is high：Concentration is in 300-2000ug/ul；Quality is high：A260/
A280 values are more than 1.8；Prove that carried RNA is suitble to quantitative fluorescent PCR and transcript profile sequencing by subsequent experimental.
Certainly, it implements any of the products of the present invention it is not absolutely required to while reaching all the above technique effect.
Description of the drawings
Attached drawing described herein is used to provide further understanding of the present invention, and constitutes the part of the present invention, this hair
Bright illustrative embodiments and their description are not constituted improper limitations of the present invention for explaining the present invention.In the accompanying drawings：
Fig. 1 is the procedure chart of fish-skin stripping of the present invention and striped sample collection；Wherein, a represents experiment middle Warsaw loach used；
B representatives strip skin since gill cover portion；C representatives are puted forth effort from husky loach nape part, strip monoblock skin；D representatives strip monoblock skin
It paves after skin, is fixed with liquid nitrogen；E representatives extract required parts of skin on the fixed skin of liquid nitrogen with scalpel.
Specific implementation mode
Carry out the embodiment that the present invention will be described in detail below in conjunction with embodiment, thereby to the present invention how application technology hand
Section solves technical problem and reaches the realization process of technical effect to fully understand and implement.
The foundation of the extracting method of the black yellow striped skin RNA of Warsaw loach in embodiment 1
Step 1, equipment advanced processing：The dissecting tool that will be used before extraction such as solve plane cut, scalpel, tweezers and medicine
Spoon, mortar are put into high-pressure sterilizing pot sterilizing after being wrapped up with masking foil, and are dried and are pre-chilled；
Step 2, peeling processing：
Step 2.1, operating personnel need band mask and disposable powder-free gloves, are sprayed with alcohol watering can and gloves and wipe dry, made
Replace new gloves in due course in；
Step 2.2, fish-skin stripping：As shown in Figure 1, since middle Warsaw loach is alepidote, take skin without removing scale (figure
1a)；First fish is carried out with 40mg/L MS-222 solution to anaesthetize and break neck, then pectoral fin is cut with dissecting scissors, facilitates skinning operations；
Fish maw is broken along ventrimeson, cuts fish head along gill cover edge, then fish-skin (Fig. 1 b and Fig. 1 c) is separated from neck with scalpel, is used
Tweezers clamp fish-skin, and cooperation scalpel slowly strips out complete fish-skin, rejects a small amount of musculature adhered on fish-skin, and rapid
Complete fish-skin is laid on masking foil, liquid nitrogen is used in combination to freeze (Fig. 1 d).
Step 2.3, striped sample collection：Due to fish-skin softness flexible, it is desirable to it is directly separated black yellow striped and is not easy to,
So freezed on masking foil fish-skin using liquid nitrogen, then the texture with scalpel along black yellow striped is directly cut (figure
1e)；It is operated in the case of liquid nitrogen frozen, can quickly separate the black yellow striped of fish-skin, also can guarantee that its RNA is non-degradable.
Striped skin is then packed into 2ml cryopreservation tubes, is put into liquid nitrogen.
Step 3, extraction step：Middle Warsaw loach striped skin is ground rapidly in precooling completely and the mortar equipped with liquid nitrogen
(since skin histology hardness is big, flexible, grind away difficulty is larger), side is ground at this time, while shredded skin histology with dissecting scissors,
More the thinner fritter the better, is further continued for grinding until it is in powdered；
60-80mg skin histology powder is taken to be added in the EP pipes for the Trizol liquid for having filled 1ml with the spoon of Liquid nitrogen precooler,
Fully oscillation is uniformly mixed, and is put into 12000g, and supernatant is transferred to new centrifuge tube by 4 DEG C of centrifugation 5min；It is centrifuged in previous step
1/5 volume (200ul) chloroform CHCl is added in pipe3, 15s vortex mixings, after standing 5min, 12000g, 4 DEG C of centrifugation 15min；It will
Supernatant is transferred to new centrifuge tube；It is primary to repeat previous step, preceding once to act as extracting, latter step is elution, improves RNA
Purity；It is transferred to new centrifuge tube, isometric isopropanol is added, mixing is stored at room temperature 10min, 12000g, 4 DEG C, centrifuges 10min；
Supernatant is abandoned, 1ml ice ethyl alcohol (cleaning action) is added in precipitation, 12000g, centrifuges 5min by 4 DEG C；Supernatant is abandoned, it is drying precipitated,
It is dissolved in appropriate DEPC water.
To ensure having liquid nitrogen in mortar always in process of lapping, keep material non-degradable, and as far as possible with dissecting scissors by skin
It is cut into fine particle to regrind to powder, the skin histology not ground powder and is transferred in Trizol liquid, is discarded after centrifugation
Lower layer's impurity.
Step 4, quality testing：
It is detected (principle is spectrophotometer measurement) using nucleic acid-protein instrument NanoDrop2000
Step 4.1 first draws 100＆mu；The DEPC water of l, is put into instrument, point blank keys, adjusts standard zero.
Step 4.2 after cleaning measuring appliance, draws 1ul samples, puts on measuring appliance, point measure keys measure, then
Obtain concentration results.
By experiment, the black yellow skin RNA concentration quality of middle Warsaw loach prepared with the method is high, effectively reduces routine sampling
Muscle skin caused by method is difficult to detach, grind away time and effort consuming, RNA are impure, is asked by what higher muscle protein content was influenced
Topic.Conventional method sampling difficulty, it is desirable to take clean intact skin sample difficult, especially on alepidote, skin is thin, toughness
By force, muscle skin is tightly connected and is difficult to divide, and causes to be difficult to obtain required sample to be tested.
Said extracted method, utensil are simple, it is most important that the fixed opportunity palm of gimmick and liquid nitrogen that fish-skin skin strips
It holds, by many experiments, discovery is operated on from fish belly portion, is cut to neck along the gill cover, could keep trunk striped integrality, then from neck
Portion, which takes advantage of a situation to tear toward tail portion, takes skin, can ensure that whole skin is completely stripped down.Stripping first first cuts off pectoral fin dorsal fin, in order to avoid stripping
It is obstructed during taking, tears skin.
The time of skin is fixed for liquid nitrogen, the time, few then skin still relaxed, and scalpel extracts inconvenience, and RNA can degrade, when
Between it is long, freeze too hard, cut and do not break, influence experiment process, after skin is tiled, it is best to pour liquid nitrogen frozen 30-50 seconds.
The application examples of the extracting method of the black yellow striped skin RNA of Warsaw loach in embodiment 2
In the acquisition experiment of the thin loach dermatological specimens of length, using method described in embodiment 1, long thin loach individual is larger, skin
Warsaw loach is coarse in skin ratio, is easy to cut required blotchy skin with previous method, with the method that skin is substantially complete
It is whole remove after, fixed, then directly extract the black line of spot and adjacent yellow line with scalpel, extracted after RNA for being sequenced, with liquid nitrogen
Meet requirement for construction data base, as shown in table 1.
After the extraction of table 1 RNA library index is built for sequencing
Above description has shown and described several preferred embodiments of invention, but as previously described, it should be understood that invention is not
It is confined to form disclosed herein, is not to be taken as excluding other embodiments, and can be used for various other combinations, modification
And environment, and can be carried out by the above teachings or related fields of technology or knowledge in the scope of the invention is set forth herein
Change.And changes and modifications made by those skilled in the art do not depart from the spirit and scope of invention, then should all be weighed appended by invention
In the protection domain that profit requires.
Claims (4)
1. the extracting method of the black yellow striped skin RNA of middle Warsaw loach a kind of, which is characterized in that include the following steps：Equipment is located in advance
Reason, peeling processing, extraction step, quality testing.
2. extracting method according to claim 1, which is characterized in that the equipment in the step 1, which pre-processes, is specially：It carries
The dissecting tool that will be used before taking such as solves after plane is cut, scalpel, tweezers and spoon, mortar are wrapped up with masking foil and is put into high pressure
Autoclave sterilizes, and is dried and is pre-chilled.
3. extracting method according to claim 1, which is characterized in that the peeling in the step 2, which is handled, is specially：
Step 2.1, operating personnel need band mask and disposable powder-free gloves, with alcohol watering can spray gloves and wipe it is dry, in use
New gloves are replaced in due course；
Step 2.2, fish-skin stripping：Since middle Warsaw loach is alepidote, take skin without removing scale；First use 40mg/L MS-222
Solution carries out fish to anaesthetize and break neck, then cuts pectoral fin with dissecting scissors, facilitates skinning operations；Fish maw is broken along ventrimeson, along the gill
Fish head is cut at lid edge, then separates fish-skin from neck with scalpel, clamps fish-skin with tweezers, cooperation scalpel slowly will be complete
Fish-skin strips out, and rejects a small amount of musculature adhered on fish-skin, and complete fish-skin is laid on masking foil rapidly, liquid nitrogen is used in combination
It freezes；
Step 2.3, striped sample collection：Due to fish-skin softness flexible, it is desirable to it is directly separated black yellow striped and is not easy to, so
Fish-skin is freezed on masking foil using liquid nitrogen, then the texture with scalpel along black yellow striped is directly cut；In liquid nitrogen
It is operated in the case of freezing, can quickly separate the black yellow striped of fish-skin, also can guarantee that its RNA is non-degradable.Then by striped
Skin is packed into 2ml cryopreservation tubes, is put into liquid nitrogen.
4. extracting method according to claim 1, which is characterized in that the extraction step in the step 3 is specially：Will in
Warsaw loach striped skin is pre-chilled and is ground rapidly in mortar equipped with liquid nitrogen complete, and side is ground at this time, while with dissecting scissors by skin
Skin tissue shreds, and more the thinner fritter the better, is further continued for grinding until it is in powdered；60-80mg is taken with the spoon of Liquid nitrogen precooler
Skin histology powder is added in the EP pipes for the Trizol liquid for having filled 1ml, and fully oscillation is uniformly mixed, and is put into 12000g, 4 DEG C from
Supernatant is transferred to new centrifuge tube by heart 5min；1/5 volume of chloroform CHCl is added in previous step centrifuge tube3, 15s vortexs
Mixing, after standing 5min, 12000g, 4 DEG C of centrifugation 15min；Supernatant is transferred to new centrifuge tube；Repeat previous step one
Secondary, preceding once to act as extracting, latter step is elution, improves RNA purity；It is transferred to new centrifuge tube, isometric isopropanol is added,
Mixing is stored at room temperature 10min, 12000g, 4 DEG C, centrifuges 10min；It abandons supernatant, is added 1ml ice ethyl alcohol in precipitation, 12000g, 4
DEG C, centrifuge 5min；Supernatant is abandoned, it is drying precipitated, it is dissolved in appropriate DEPC water.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020091044A1 (en) * | 2018-11-01 | 2020-05-07 | 花王株式会社 | Method for preparing nucleic acid derived from skin cell of subject |
| CN114397158A (en) * | 2022-01-07 | 2022-04-26 | 重庆师范大学 | Preparation method of fish early warning pheromone |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060093596A1 (en) * | 2002-08-22 | 2006-05-04 | Susan Douglas | Genomic approach to identification of novel broad-spectrum antimicrobial peptides from bony fish |
| CN102559695A (en) * | 2011-11-16 | 2012-07-11 | 集美大学 | Pseudosciaena crocea interferon (IFN) gene and cloning method thereof |
| CN103113461A (en) * | 2012-11-06 | 2013-05-22 | 中国农业科学院饲料研究所 | Novel defensin BD2, and gene and application thereof |
| CN103160497A (en) * | 2013-04-07 | 2013-06-19 | 北京市水产科学研究所 | Method for rapidly extracting total RNA (ribonucleic acid) from fish |
| CN104830840A (en) * | 2015-05-26 | 2015-08-12 | 中国水产科学研究院黄海水产研究所 | Pretreatment method for cynoglossus semilaevis skin tissue RNA extraction |
| CN105624322A (en) * | 2016-03-30 | 2016-06-01 | 集美大学 | Method for developing SSR (Simple Sequence Repeat) molecular mark of nibea albiflora for population identification |
| CN106636204A (en) * | 2017-01-09 | 2017-05-10 | 华中农业大学 | Breeding method of albino paramisgurnus dabryanus capable of being inherited stably |
| CN107574168A (en) * | 2017-11-02 | 2018-01-12 | 中国水产科学研究院淡水渔业研究中心 | A kind of miRNA of regulation and control red tilapia body-color diversity and its application |
-
2018
- 2018-05-08 CN CN201810432740.0A patent/CN108611347A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20060093596A1 (en) * | 2002-08-22 | 2006-05-04 | Susan Douglas | Genomic approach to identification of novel broad-spectrum antimicrobial peptides from bony fish |
| CN102559695A (en) * | 2011-11-16 | 2012-07-11 | 集美大学 | Pseudosciaena crocea interferon (IFN) gene and cloning method thereof |
| CN103113461A (en) * | 2012-11-06 | 2013-05-22 | 中国农业科学院饲料研究所 | Novel defensin BD2, and gene and application thereof |
| CN103160497A (en) * | 2013-04-07 | 2013-06-19 | 北京市水产科学研究所 | Method for rapidly extracting total RNA (ribonucleic acid) from fish |
| CN104830840A (en) * | 2015-05-26 | 2015-08-12 | 中国水产科学研究院黄海水产研究所 | Pretreatment method for cynoglossus semilaevis skin tissue RNA extraction |
| CN105624322A (en) * | 2016-03-30 | 2016-06-01 | 集美大学 | Method for developing SSR (Simple Sequence Repeat) molecular mark of nibea albiflora for population identification |
| CN106636204A (en) * | 2017-01-09 | 2017-05-10 | 华中农业大学 | Breeding method of albino paramisgurnus dabryanus capable of being inherited stably |
| CN107574168A (en) * | 2017-11-02 | 2018-01-12 | 中国水产科学研究院淡水渔业研究中心 | A kind of miRNA of regulation and control red tilapia body-color diversity and its application |
Non-Patent Citations (3)
| Title |
|---|
| WENBIN ZHU等: "Comparative Transcriptome Analysis Identifies Candidate Genes Related to Skin Color Differentiation in Red Tilapia", 《SCI REP.》 * |
| 徐丹丹等: "中华沙鳅和宽体沙鳅形态差异及其物种有效性分析", 《水生生物学报》 * |
| 覃川杰等: "宽体沙鳅（Botia reevesae）β-肌动蛋白基因的cDNA克隆与表达分析", 《海洋与湖沼》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020091044A1 (en) * | 2018-11-01 | 2020-05-07 | 花王株式会社 | Method for preparing nucleic acid derived from skin cell of subject |
| JP2020074769A (en) * | 2018-11-01 | 2020-05-21 | 花王株式会社 | Method for preparing nucleic acid derived from skin cells of subject |
| CN114397158A (en) * | 2022-01-07 | 2022-04-26 | 重庆师范大学 | Preparation method of fish early warning pheromone |
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Application publication date: 20181002 |