CN110396541A - Schizophrenia disease mouse model hippocampus lncRNA sequencing analysis and kit - Google Patents
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Abstract
The invention belongs to field of biotechnology, are related to schizophrenia neurodevelopment mouse model hippocampus biomarker, detection method and kit.The present invention provides: a kind of 1. schizophrenia neurodevelopment mouse model hippocampus biomarker, for lncRNA2, lncRNA4, lncRNA5, lncRNA6, the detection method of above-mentioned marker and the kit of the above-mentioned marker content of detection；It is lncRNA6, the detection method of above-mentioned marker and the kit of the above-mentioned marker content of detection 2. additionally providing a kind of biomarker that schizophrenia neurodevelopment mouse model hippocampus is intervened；3. providing the GO/KEGG enrichment analysis result and lncRNA-TF relational network figure of differential expression lncRNA6 in hippocampus.The beneficial effects of the present invention are stress stimulations after having proved the nervous lesion of mouse pregnancy period, birth can cause schizophrenia original mold type, and find that its hippocampus lncRNA has expression otherness for the first time, the variation can be reversed by Olanzapine intervention, this is to the relationship of schizophrenic onset and epigenetics and finds its new drug and is of great significance.
Description
Technical field
The present invention relates to field of biotechnology, especially a kind of schizophrenia disease mouse model hippocampus lncRNA sequencing analysis
And kit.
Background technique
Schizophrenia (schizophrenia, SZ) is a kind of
Main principal characteristic mental disease, pathogenesis are still not clear, and affect the population in the whole world 1%, cause great social economy negative
Load；The disease can hide onset, the course of disease is hidden, prognosis mala, recurrent exerbation, and clinical diagnosis relies primarily on semiotics, and there are subjectivities
Bias, drug therapy mainly include the second generation atypical antipsychotic including Olanzapine (olanzapine, OLZ).
With the arrival in post-genomic science epoch, transcription group is that initiative development gets up and most widely used technology.
Transcript profile is the summation for all RNA that specific organization or cell transcribe out under a certain stage of development or functional status, mainly
Including mRNA and non-coding RNA (non-coding RNA, ncRNA).Researcher's recent years discovery, long-chain non-coding RNA
(long non-coding RNA, lncRNA) plays regulating and controlling effect in mental disorder pathomechanism.LncRNA was in 2002
It is found for the first time, is the RNA molecule that a kind of Transcript Length is more than 200nt, in vivo wide participation DNA methylation, group egg
It is white modification, chromosome remodeling etc. biological processes, in the form of RNA in a variety of levels (epigenetic regulation, transcriptional control with
And post-transcriptional control etc.) controlling gene expression, as X chromosome silencing, genomic imprinting and chromatin modification, turn
Record, montage, translation, intracellular transport etc., it is closely related with the generation, development, diagnosing and treating of a variety of diseases.
Schizophrenia is neurodevelopment sexual dysfunction, is assessed cerebral tissue lncRNA expression and intervention effect advantageous
In further regulatory mechanism and therapy target of the announcement lncRNA in schizophrenia.But it is limited by Bioethics
Limitation, the acquisition based on human schizophrenia living body brain tissue are difficult to carry out.Recently, schizophrenia neurodevelopment mouse mould
Type is had mature theoretical basis, modeling standard and Testing index by researcher's attention at present, in mechanism validity, apparently
There is apparent advantage in terms of hereditary validity and predictive validity.Therefore, the invention is intended to inquire into schizophrenia neurodevelopment
Mouse model hippocampus lncRNA expression, detection method and kit.
Summary of the invention
In order to overcome deficiency existing for existing technology, the present invention provides a kind of schizophrenia disease mouse model hippocampus
LncRNA sequencing analysis and kit.
The technical solution adopted by the present invention to solve the technical problems is: a kind of schizophrenia neurodevelopment mouse model
Hippocampus biomarker, marker lncRNA2, lncRNA4, lncRNA5, lncRNA6.
A kind of detection method of schizophrenia neurodevelopment mouse model hippocampus biomarker, comprising the following steps:
Brain tissue acquisition, three groups of control group (NC), schizophrenia model group (SZ), treatment group (Treated) mouse are equal
In 100 days sacrificed by decapitation and remove full brain；
Library extraction sample total serum IgE is built in RNA extracting, and after use kit ribo-zero digestion rRNA；Addition is beaten
RNA is broken into short-movie section by disconnected reagent, using the RNA after interrupting as template, synthesizes a chain cDNA with hexabasic base random primer, then
It prepares two chain synthesis reaction systems and synthesizes two chain cDNA, dTTP is replaced with dUTP in the synthesis of bis- chain of cDNA, then connection is different
Connector recycles UNG enzyme process to digest a chain containing dUTP, only retains mono- chain of cDNA of connection chain difference connector；
Use mono- chain of kits cDNA；Mono- chain of cDNA of purifying carries out end reparation plus A tail again and connects sequence measuring joints, then
Clip size selection is carried out, PCR amplification is finally carried out；
After the 2100 Bioanalyzer quality inspection qualification of Agilent of the library built, Illumina HiSeqTM is used
2500 or other sequencers；Real-Time PCR (RT-PCR) verifying, according to sequencing result and standard rna sequence, design
LncRNA probe and primer；It can be detected according in fold differences FC >=2, cluster analysis result in different clusters, each sample standard deviation
Three conditions select 10 difference lncRNA of hippocampus of mice respectively and carry out RT-RCR detection out；Each lncRNA sample repeats in parallel
3 times；Whether the differential expression for comparing NC group and SZ group is significant, chooses the lncRNA that there were significant differences again in SZ group and Treated
RT-PCR detection is carried out in group sample.
A kind of schizophrenia neurodevelopment mouse model hippocampus biomarker kit, can measure lncRNA2,
LncRNA4, lncRNA5, lncRNA6 expression.
A kind of biomarker that schizophrenia neurodevelopment mouse model lapses to, the marker are lncRNA6.
A kind of schizophrenia neurodevelopment mouse model lapses to the detection method of biomarker,
The hippocampal tissue of schizophrenia neurodevelopment mouse model after taking appropriate antipsychotic drug (Olanzapine) to treat, from
Middle extraction total serum IgE, and reverse transcription reaction is carried out after carrying out normal state correction；Real-time fluorescence quantitative PCR is carried out after the reaction was completed, is obtained
To lncRNA6 expression；
To select that internal reference can be tested as RT-PCR using the β-Actin that stable detection arrives in all samples by test,
The relative expression levels of target RNA are indicated with Δ Ct value (Δ Ct=Cttarget-Ct β-actin)；Use FPKM method
(Fragments Per kb Per Million Reads) counts target rna expression amount；Using NB (negative binomial distribution inspection)
The test-target RNA significance of difference；
By establishing the norm of lncRNA6 expression, standardized value is obtained.
A kind of schizophrenia neurodevelopment mouse model lapses to detection kit, can measure in hippocampal tissue
The content of lncRNA6.
Difference lncRNA GO/KEGG enrichment analysis result in a kind of offer lncRNA-TF relational network figure and hippocampus.
The beneficial effects of the present invention are:
1, the building of schizophrenia neurodevelopment mouse model will increase new objective Testing index, can by pair
LncRNA detection, improves accuracy, standardization and the success rate of schizophrenia neurodevelopment mouse model；
2, it is expected to change specific lncRNA expression energy by lncRNA analogies and antagonist (modified oligonucleotide)
Enough make abnormal gene regulatory network and signal path more normalization, and then treats schizophrenia；
3, it is expected to predict medication curative effect by detecting specific lncRNA expression；
4, it is expected to lapse to schizophreniac by the specific lncRNA expression of detection and prognosis situation carries out in advance
It surveys；
5, it is expected to carry out specific aim to certain specific symptoms of specific schizophrenia by lncRNA analogies and antagonist
Treatment.
Detailed description of the invention
Present invention will be further explained below with reference to the attached drawings and examples.
The dendrogram of Figure 1A expression full transcriptome analysis of model group hippocampus difference RNA；Figure 1B indicates medication group hippocampus difference
The dendrogram (screening conditions: P<0.05 and fold differences>2) of the full transcriptome analysis verifying of RNA, red represent relatively high expression,
Green represents opposite low expression.
Fig. 2A is the PCR verifying of differential expression lncRNA expression in each group hippocampus of mice；Fig. 2 B is medication group mouse
The PCR verifying of differential expression lncRNA in hippocampus.
Fig. 3 A is lncRNA-TF binary crelation network；Fig. 3 B is lncRNA-TF ternary relation network.
Specific embodiment
A kind of schizophrenia neurodevelopment mouse model hippocampus biomarker, marker lncRNA2, lncRNA4,
lncRNA5、lncRNA6。
A kind of detection method of schizophrenia neurodevelopment mouse model hippocampus biomarker, comprising the following steps:
Brain tissue acquisition, three groups of control group (NC), schizophrenia model group (SZ), treatment group (Treated) mouse are equal
In 100 days sacrificed by decapitation and remove full brain；
Library extraction sample total serum IgE is built in RNA extracting, and after use kit ribo-zero digestion rRNA；Addition is beaten
RNA is broken into short-movie section by disconnected reagent, using the RNA after interrupting as template, synthesizes a chain cDNA with hexabasic base random primer, then
It prepares two chain synthesis reaction systems and synthesizes two chain cDNA, dTTP is replaced with dUTP in the synthesis of bis- chain of cDNA, then connection is different
Connector recycles UNG enzyme process to digest a chain containing dUTP, only retains mono- chain of cDNA of connection chain difference connector；
Use mono- chain of kits cDNA；Mono- chain of cDNA of purifying carries out end reparation plus A tail again and connects sequence measuring joints, then
Clip size selection is carried out, PCR amplification is finally carried out；
After the 2100 Bioanalyzer quality inspection qualification of Agilent of the library built, Illumina HiSeqTM is used
2500 or other sequencers；Real-Time PCR (RT-PCR) verifying, according to sequencing result and standard rna sequence, design
LncRNA probe and primer；It can be detected according in fold differences FC >=2, cluster analysis result in different clusters, each sample standard deviation
Three conditions select 10 difference lncRNA of hippocampus of mice respectively and carry out RT-RCR detection out；Each lncRNA sample repeats in parallel
3 times；Whether the differential expression for comparing NC group and SZ group is significant, chooses the lncRNA that there were significant differences again in SZ group and Treated
RT-PCR detection is carried out in group sample.
A kind of schizophrenia neurodevelopment mouse model hippocampus biomarker kit, can measure lncRNA2,
LncRNA4, lncRNA5, lncRNA6 expression.
A kind of biomarker that schizophrenia neurodevelopment mouse model lapses to, the marker are lncRNA6.
A kind of schizophrenia neurodevelopment mouse model lapses to the detection method of biomarker,
The hippocampal tissue of schizophrenia neurodevelopment mouse model after taking appropriate antipsychotic drug (Olanzapine) to treat, from
Middle extraction total serum IgE, and reverse transcription reaction is carried out after carrying out normal state correction；Real-time fluorescence quantitative PCR is carried out after the reaction was completed, is obtained
To lncRNA6 expression；
To select that internal reference can be tested as RT-PCR using the β-Actin that stable detection arrives in all samples by test,
The relative expression levels of target RNA are indicated with Δ Ct value (Δ Ct=Cttarget-Ct β-actin)；Use FPKM method
(Fragments Per kb Per Million Reads) counts target rna expression amount；Using NB (negative binomial distribution inspection)
The test-target RNA significance of difference；
By establishing the norm of lncRNA6 expression, standardized value is obtained.
A kind of schizophrenia neurodevelopment mouse model lapses to detection kit, can measure in hippocampal tissue
The content of lncRNA6.
Difference lncRNA GO/KEGG enrichment analysis result in a kind of offer lncRNA-TF relational network figure and hippocampus.
1, research object
This research uses C57BL/6 mouse, is often considered as the inbred mouse of " standard ", the heredity back with height
Scape stability is the mammal of first completion gene order-checking after the mankind.7~8 week old C57BL/6 mouse of parental generation,
It is bought, SPF grades from Jiangsu University's Experimental Animal Center (experimental animal production licence number: SCXK (Soviet Union) 2013-0011), it is male
Property 20, female 45 continues 2 Zhou Houkai of raising by male and female, with every 4~5 sub-cage rearings of gender in new animal feeding room
The breeding of beginning offspring rat.21 ± 1 DEG C of animal feeding room temperature, illumination round the clock overturn (turn off the light: 08:00~20:00, the 20:00 that turns on light~
08:00), unless otherwise specified, all animals have sufficient food and water for its free feeding.
Offspring rat breeding: to make female mice oestrus unanimously to obtain the consistent pregnant mouse of period of pregnancy, according to Meyer etc.
Report, separated by every 2~3 female mices with 1 male mouse wire netting within 3 days before mating and mate raising, in 08:00 removal in the 4th day
Wire netting, every 4~5h check primary mating situation, find vaginal plug and be denoted as pregnant 0 day (GD0).More than 5 days ends female mice of becoming pregnant is given
It excludes.
This research obtains the approval of the 904th Animal Experimental Ethical committee, hospital of liberation army.
2, research method
1. parent pregnancy period immunostimulation: immunostimulation group (P+) mouse is infused in female mice GD9 by 6mg/kg dosage tail vein
Penetrate (i.v.) Poly (I:C), Poly (I:C) solution is prepared with sterile saline, and is diluted to 1.2mg/ml, finally with
The volume of 5ml/kg is injected, and nonimmune stimulation group (P-) then injects sterile saline.All injection solution are existing with existing
With.
2. offspring rat puberty stress: through Poly (I:C) or physiological saline treated the produced offspring rat of female mice in
Weaned and divided within the 21st day (PND21) after birth cage (rearging cage specification: 28cm × 18cm × 14cm), to reduce litter effect
Influence, littermate by gender is evenly distributed to puberty stress group (S+) and without in stress group (S-).Offspring rat presses gender
And processing factor, reason factor is existed together by the raising of every 3-4/cage with gender.Puberty stress group carries out chronic in PND31~40 day
It is unknowable to intervene, then normally raising not other any intervention processing of no stress group.With reference to Ducottet's and Sandra
Design, puberty is chronic unknowable stress successively to be given as follows: soaking sawdust padding (Day1), rearging cage tilts 45 °
(Day2), foot shock (Day3), food deprivation (Day4), without padding (Day5), Restraint Stress (Day6), water deprivation
(Day7), night mutually illuminate (Day8), forced swimming (Day9), social activity stress (Day10).
3. Olanzapine (Olanzapine, OLZ) standard items used in this research of pharmaceutical intervention are ground by the calibrating of Chinese instrumented medication
Study carefully institute's (ID:V4V2069GU, number: 100948-200801) to provide.By receive it is dual stress mouse be divided into medication group
(Treated) and model group (SZ) the medication group mouse for, receiving OLZ intervention, is given by the daily drinking-water of mouse, dosage
1.5mg/ (kgd), drinking bottle avoid medical fluid from decomposing because of illumination by being protected from light processing.The administration same day measures mouse weight and preceding 4
It drinking-water consumption calculates the drinking-water concentration of first administration.After OLZ is dissolved with a small amount of glacial acetic acid, it is diluted to distilled water
1mg/ml adjusts PH to 6.0 ± 0.2 with NaHCO3, calculates drinking water administration concentration according to every cage mouse weight and drinking-water consumption,
Prepared medical fluid is diluted to aimed concn.A weight and drinking-water consumption are re-measured within the every cage of medication group mouse every 4 days,
Correction drinking-water drug concentration, and replace drinking-water medical fluid.Model group then normal water.
4. prepulse inhibition (PPI) tests this research and is startled reflected P PI test macro (SR-LAB, San Diego by sound
Instruments, San Diego, CA, USA) PPI function is checked.Entire test process is made of 4 kinds of stimulus types:
Single Pulse (white noise of 120dBA, duration 40ms), single Prepulse are (not comprising 5 kinds of 69,73,77,81 and 85dBA
With the white noise of sound pressure level, i.e., respectively higher than 4,8,12,16 and 20dBA of background noise, continue 20ms), Prepulse+Pulse
(Prepulse and Pusle start the time interval of output as 100ms) and Background (only background noise 65dBA).Entirely
Stimulus sequence includes 6 initial Pulse, intermediate 12 Block (Pulse × 1, Prepulse × 5, Prepulse+
Pulse × 5 and Background × 1, pseudorandom permutation arrangement processing) and end up 6 Pulse.Sonic stimulation sequence adjacent two
15 ± 5s (as 10~20s) is divided between a stimulation.Evaluation index: corresponding to each varying strength Prepulse+Pulse,
PPI function indicates with prepulse inhibition rate (PPI%), formula are as follows: PPI%=[(Pulse average response intensity-Prepulse+
Pulse average response intensity)/Pulse average response intensity] × 100%.Wherein, each 6 Pulse of beginning and end are not included in
It calculates.
5. brain tissue acquires three groups of control group (NC), schizophrenia (SZ), treatment group (Treated) mouse in 100
Its sacrificed by decapitation simultaneously removes full brain, and the mouse brain outside of belly is put into the mouse brain coronal section mold (037- of ice bag pre-cooling upward
001, HuaYueYang Biotechnology, Beijing, China), Mouse Whole Brain is cut into the hat of 1mm thickness with double-edged razor blade
The blade for being stained with brain hat load is laid on ice bag by shape slice.Control mice brain anatomical atlas, is looked for referring to the position Bregma
To AP (- 1.3~-3.3) (including hippocampus), coronal target area brain tissue of brain is taken with diameter 1mm flat mouth syringe needle stamp, immerses dress
Have 10ul RNA tissue preserration liquid RNA guarder (ZH0103-A, HuaYueYang Biotechnology, Beijing,
China in 1.5ml centrifuge tube), -80 DEG C of refrigerators of immigration are frozen spare afterwards.
6. library extraction sample total serum IgE is built in RNA extracting, and after use kit ribo-zero digestion rRNA；It is added
It interrupts reagent and RNA is broken into short-movie section, using the RNA after interrupting as template, synthesize a chain cDNA with hexabasic base random primer, so
After prepare two chain synthesis reaction systems synthesize two chain cDNA, bis- chain of cDNA synthesis when with dUTP replace dTTP, then connect not
Same connector recycles UNG enzyme process to digest a chain containing dUTP, only retains the cDNA mono- of connection chain difference connector
Chain；Use mono- chain of kits cDNA；Mono- chain of cDNA of purifying carries out end reparation plus A tail again and connects sequence measuring joints, so
Clip size selection is carried out afterwards, finally carries out PCR amplification；The library built 2100 Bioanalyzer quality inspection of Agilent
After qualification, Illumina HiSeqTM 2500 or other sequencers are used.
7. Real-Time PCR (RT-PCR) verifying according to sequencing result and standard rna sequence, design lncRNA probe and
Primer.Three conditions point can be detected in different clusters, each sample standard deviation according in fold differences FC >=2, cluster analysis result
10 difference lncRNA of hippocampus of mice are not selected carries out RT-RCR detection.Each lncRNA sample is repeated 3 times in parallel.Compare NC group
It is whether significant with the differential expression of SZ group, it chooses the lncRNA that there were significant differences and is carried out in SZ group and Treated group sample again
RT-PCR detection.
Total data carries out statistical using Graph Pad Prism 7.0 and 5.0 software of StatView Version
Analysis.It selects that internal reference can be tested as RT-PCR using the β-Actin that stable detection arrives in all samples by test.With Δ Ct value
The relative expression levels of (Δ Ct=Cttarget-Ct β-actin) expression target RNA.Use FPKM method (Fragments Per
Kb Per Million Reads) statistics target rna expression amount.Using NB (negative binomial distribution inspection) test-target RNA difference
Conspicuousness.Using between independent sample T inspection, one-way analysis of variance, non-parametric test Kruscal-Wallis Test analysis group
Target RNA Δ Ct value difference is different.Differential transcription combination of sets is using GO functional analysis and KEGG path analysis.Using Pearson method into
Correlation analysis between row difference RNA.All statistical analysis are two-sided criterion of significance, are that difference has statistics with P < 0.05
Meaning.
3, result of study
1. the lncRNA analysis of differential expression in each group hippocampus of mice
Compared with the control group, there are the lncRNA that 598 species diversity are expressed for model group, wherein 249 kinds of up-regulations, 349 kinds of downwards.
See attached drawing 1A.
Medication group and model group comparison find that there are 820 differential expressions by lncRNA, wherein 427 are raised, under 393
It adjusts.See attached drawing 1B.
2. the RT-PCR verifying of difference RNA in each group hippocampus of mice
LncRNA2, lncRNA4, lncRNA5, lncRNA6 expression are lowered in model group, and equal difference has statistics meaning
Justice is shown in attached drawing 2A.LncRNA6 expression raises in medication group, and difference is statistically significant, sees attached drawing 2B.
3. the functional analysis of lncRNA in model group
GO/KEGG enrichment analysis is carried out to lncRNA2, lncRNA4, lncRNA5, lncRNA6 of differential expression in hippocampus,
It is as shown in table 1:
The GO/KEGG of differential expression IncRNA is enriched with analysis in 1 hippocampus of table
Note: ID: number；Fold-Change: fold differences；P-value:P value；GO/KEGG-description:-GO/
The description of KEGG entry.
4. lncRNA-TF relational network figure constructs
Fig. 3 A is lncRNA-TF binary crelation network；Fig. 3 B is lncRNA-TF ternary relation network.Blue node
Transcription factor is represented, red node indicates LncRNA, and green node indicates target gene；Point size is wired to outward with it
Direct ratio, relationship is directly proportional to the number of appearance in the thickness and statistical result of line；Nodename is located at immediately below node figure.
In summary, it is believed that lncRNA6 can be as reflection schizophrenia neurodevelopment mouse model biology mark
Remember object.
All documents that the present invention refers to are incorporated herein by reference, and are individually drawn just as each document
It is used as with reference to such.
Described above to be merely exemplary for the purpose of the present invention, and not restrictive, those of ordinary skill in the art understand,
In the case where not departing from spirit and scope as defined in the appended claims, many modifications, variation or equivalent can be made, but all
It will fall within the scope of protection of the present invention.
Claims (7)
1. a kind of schizophrenia neurodevelopment mouse model hippocampus biomarker, characterized in that marker lncRNA2,
lncRNA4、lncRNA5、lncRNA6。
2. a kind of detection side of schizophrenia neurodevelopment mouse model hippocampus biomarker according to claim 1
Method, characterized in that the following steps are included:
Brain tissue acquisition, three groups of control group (NC), schizophrenia model group (SZ), treatment group (Treated) mouse are in 100
Its sacrificed by decapitation simultaneously removes full brain；
Library extraction sample total serum IgE is built in RNA extracting, and after use kit ribo-zero digestion rRNA；Addition interrupts examination
RNA is broken into short-movie section by agent, using the RNA after interrupting as template, is synthesized a chain cDNA with hexabasic base random primer, is then prepared
Two chain synthesis reaction systems synthesize two chain cDNA, replace dTTP in the synthesis of bis- chain of cDNA with dUTP, then connect different connectors,
It recycles UNG enzyme process to digest a chain containing dUTP, only retains mono- chain of cDNA of connection chain difference connector；Use examination
Agent box purifies mono- chain of cDNA；Mono- chain of cDNA of purifying carries out end reparation plus A tail again and connects sequence measuring joints, then carries out piece
Duan great little selection, finally carries out PCR amplification；
After the 2100 Bioanalyzer quality inspection qualification of Agilent of the library built, Illumina HiSeqTM is used
2500 or other sequencers；Real-Time PCR (RT-PCR) verifying, according to sequencing result and standard rna sequence, design
LncRNA probe and primer；It can be detected according in fold differences FC >=2, cluster analysis result in different clusters, each sample standard deviation
Three conditions select 10 difference lncRNA of hippocampus of mice respectively and carry out RT-RCR detection out；Each lncRNA sample repeats in parallel
3 times；Whether the differential expression for comparing NC group and SZ group is significant, chooses the lncRNA that there were significant differences again in SZ group and Treated
RT-PCR detection is carried out in group sample.
3. a kind of schizophrenia neurodevelopment mouse model hippocampus biomarker kit according to claim 1,
It is characterized in that lncRNA2, lncRNA4, lncRNA5, lncRNA6 expression can be measured.
4. a kind of biomarker that schizophrenia neurodevelopment mouse model lapses to, characterized in that the marker is
lncRNA6。
5. the detection side that a kind of schizophrenia neurodevelopment mouse model according to claim 4 lapses to biomarker
Method, characterized in that
The hippocampal tissue of schizophrenia neurodevelopment mouse model, Cong Zhongti after taking appropriate antipsychotic drug (Olanzapine) to treat
Total serum IgE is taken, and carries out reverse transcription reaction after carrying out normal state correction；Real-time fluorescence quantitative PCR is carried out after the reaction was completed, is obtained
LncRNA6 expression；
To select that internal reference can be tested as RT-PCR using the β-Actin that stable detection arrives in all samples by test, with Δ
The relative expression levels of Ct value (Δ Ct=Cttarget-Ct β-actin) expression target RNA；Use FPKM method (Fragments
Per kb Per Million Reads) statistics target rna expression amount；Using NB (negative binomial distribution inspection) test-target RNA
The significance of difference；
By establishing the norm of lncRNA6 expression, standardized value is obtained.
6. a kind of schizophrenia neurodevelopment mouse model according to claim 4 lapses to detection kit, feature
It is that can measure the content of lncRNA6 in hippocampal tissue.
7. difference lncRNA GO/KEGG enrichment analysis result in a kind of offer lncRNA-TF relational network figure and hippocampus.
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