CN110066872A - LncRNA UCA1 is as the application in ovarian cancer diagnosis or the biomarker of outcome inspection - Google Patents
LncRNA UCA1 is as the application in ovarian cancer diagnosis or the biomarker of outcome inspection Download PDFInfo
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Abstract
The invention discloses lncRNA UCA1 as the application in ovarian cancer diagnosis or the biomarker of outcome inspection.The application research discovery, expression of the lncRNA UCA1 in ovarian cancer tissue and cancer beside organism has significant otherness, simultaneously, research finds very likely there is interaction between lncRNA UCA1 and ovary cancer-associated protein, and the expression by adjusting ovary cancer-associated protein influences the progress of oophoroma, therefore lncRNA UCA1 can assist whether doctor suffers from oophoroma to patient or stages of ovarian carcinoma judges as the biomarker of diagnosis of ovarian cancer；While can be used as the biomarker of outcome inspection after treatment of ovarian cancer, auxiliary doctor examines the outcome of associated treatment means；LncRNA UCA1 can provide accurate believable inspection result in conjunction with other clinical indices for diagnosis, the prognosis of oophoroma.
Description
Technical field
The invention belongs to oncomolecularbiology fields, and in particular to a kind of lncRNA UCA1 is as ovarian cancer diagnosis
Or the application in the biomarker of outcome inspection.
Background technique
Oophoroma is a kind of common female malignant, and death rate of the onset has been more than cervical carcinoma and carcinoma of endometrium.
But since the early symptom of oophoroma is unobvious and more difficult detection, Most patients ovarian neoplasm when being found has existed by stages
After III phase or III phase, even across prolonged chemotherapy and operation, five year survival rate is still lower, therefore develops one kind
Specificity and the preferable cancer markers of sensitivity are found to have very important directive significance to ovarian neoplasm.
The research of lncRNAs (long non-coding RNAs, long-chain non-coding RNA) becomes in many cancers at present
Hot spot is originally found, and lncRNAs will appear unconventionality expression in the occurrence and development of many cancers: the expression of some lncRNAs is such as
It is same to be released from great expression control, and the expression of some lncRNAs seems to be inhibited by the same expression quantity to lower.The hair
Now imply that lncRNAs may be the participant during tumor development, many experimental results are also shown later
LncRNAs has the ability for promoting or inhibiting tumor development and progress.Work based on lncRNAs in terms of controlling gene expression
With and lncRNAs very likely take part in tumour, lncRNAs is deemed likely to be a kind of promising based on tissue
Or the biomarker for cancer of blood, the new paragon for the treatment of of cancer will be developed on this basis.
Such as application No. is the Chinese invention patent applications of CN201710657458.8 to disclose a kind of long-chain non-coding RNA
Application of the siRNA of SMAD5-AS1 in treatment of ovarian cancer, this application find SMAD5-AS1 in the cancerous tissue of ovarian cancer patients
In expression significantly raise, and then develop and be able to suppress in SKOV3 cell SMAD5-AS1 expression to inhibit oophoroma
The siRNA of cell progression.
The occurrence and development mechanism of cancer is still not very clear at present, and the treatment method of cancer also needs further to be developed,
And more approach can be provided for treatment of cancer by finding more cancer biomarkers objects.
Summary of the invention
Present invention purpose is to provide a kind of lncRNA UCA1 as ovarian cancer diagnosis or outcome inspection
Application in biomarker.
For achieving the above object, the technical solution of the application is as follows:
A kind of lncRNA UCA1 is as the application in ovarian cancer diagnosis or the biomarker of outcome inspection, institute
The nucleotide sequence of the lncRNA UCA1 stated is as shown in SEQ ID No.1；
Alternatively, the nucleotide sequence of the lncRNA UCA1 has with nucleotide sequence shown in SEQ ID No.1
90% or more homology.
The application is the study found that expression of the lncRNA UCA1 in ovarian cancer tissue and cancer beside organism has significantly
Otherness, meanwhile, with RPIseq line computation lncRNA UCA1 and ovary cancer-associated protein (p53, ER, FGFR-1 and
BRCA1 etc.) between when potentially interacting, find the associated score between lncRNA UCA1 and each ovary cancer-associated protein
Above 0.5, and SVM prediction score is 0.95 or so, showing between lncRNA UCA1 and ovary cancer-associated protein that pole has can
There can be interaction, and the expression by adjusting ovary cancer-associated protein influences the progress of oophoroma, therefore lncRNA
UCA1 can as the biomarker of diagnosis of ovarian cancer, assist doctor to patient whether suffer from oophoroma or stages of ovarian carcinoma into
Row judgement；It can be used as the biomarker that outcome is examined after treatment of ovarian cancer simultaneously, auxiliary doctor is to taking treatment
The progress of oophoroma is tested after means, the outcome of associated treatment means is examined, so that the treatment for oophoroma provides
New approaches and new way；LncRNA UCA1 can provide standard in conjunction with other clinical indices for diagnosis, the prognosis of oophoroma
True believable inspection result.
A kind of application of lncRNA UCA1 in the biomarker as ovarian cancer prognosis validity check, it is described
The nucleotide sequence of lncRNA UCA1 is as shown in SEQ ID No.1；
Nucleotide sequence shown in the nucleotide sequence of the lncRNA UCA1 and SEQ ID No.1 have 90% with
On homology.
Outcome examines present invention also provides a kind of for detecting the PCR primer of lncRNA UCA1 expression, should
PCR primer includes:
UCA1 upstream primer: 5 '-TCAATCAACCCTGTGACATTCTTCTCCTG-3 '；
UCA1 downstream primer: 5 '-GGAAGATTTCTTTTCTGTCACCTTTTCACATAT-3 '.
Present invention also provides a kind of for detecting the PCR kit of lncRNA UCA1 expression, the PCR kit
In containing as claimed in claim 5 for detecting the PCR primer of lncRNA UCA1 expression, and for detecting internal reference
The PCR primer of gene expression dose.
In the above-mentioned PCR kit for detecting lncRNA UCA1 expression, the reference gene is
GAPDH gene, the PCR primer for detecting reference gene expression include:
GAPDH upstream primer: 5 '-AACGGATTTGGTCGTATTG-3 '；
GAPDH downstream primer: 5 '-GGAAGATGGTGATGGGATT-3 '.
In the above-mentioned PCR kit for detecting lncRNA UCA1 expression, lncRNA UCA1's is real-time glimmering
Fluorescent Quantitative PCR reaction system includes: the qPCR reaction mixture of 5 μ L, the downstream UCA1 of the UCA1 upstream primer of 0.5 μ L, 0.5 μ L
Primer, the GAPDH upstream primer of 0.5 μ L, the GAPDH downstream primer of 0.5 μ L, 1 μ L cDNA, RNase free ddH2O is supplied
To 10 μ L.
In the above-mentioned PCR kit for detecting lncRNA UCA1 expression, lncRNA UCA1's is real-time glimmering
Fluorescent Quantitative PCR response procedures are as follows:
Stage is 1.: 95 DEG C, 5min；
Stage is 2.: 40 circular responses: 95 DEG C, 10s, 60 DEG C, 30s；
Stage is 3.: 95 DEG C, 15s, 60 DEG C, 1min, 95 DEG C, 15s.
Also containing the total serum IgE for will extract in the above-mentioned PCR kit for detecting lncRNA UCA1 expression
Reverse transcription is the reverse transcription system of cDNA, and the reverse transcription system includes M-MLV (H-) reverse transcriptase, 5 × RT M-MLV slow
Fliud flushing, dNTP mixture, UCA1 downstream primer and concentration that concentration is 2 μM are the RNase inhibitor of 40U/ μ L；
Reverse transcription reaction system includes: 5 × RT M-MLV of the total serum IgE of 2.5 μ L, the UCA1 downstream primer of 0.5 μ L, 3 μ L
Buffer, the dNTP mixture of 0.25 μ L, M-MLV (H-) reverse transcriptase of 0.5 μ L, 0.25 μ L RNase inhibitor,
Described M-MLV (H-) reverse transcriptase uses after being configured to the enzyme solutions that concentration is 200U/ μ L.
The value of the PCR kit of the application is the total serum IgE by extracting patient tissue samples, by total serum IgE reverse transcription
For cDNA, the expression of simple DNA dye method detection ovarian carcinoma associated genes UCA1 is recycled, the expression is passed through
Difference degree combines various clinicopathological parameters, can be better understood by the effect of the risk or prognosis that have ovarian cancer.Therefore should
Kit puts into production practice, to distinguishing that oophoroma and clinical auxiliary detection play an important role, if can diagnose in time
Oophoroma out, patient just have early stage cure chance, clinically for be also revolution breakthrough.
Compared with prior art, the application has the beneficial effect that:
(1) the application the study found that expression of the lncRNA UCA1 in ovarian cancer tissue and cancer beside organism have it is aobvious
The otherness of work, meanwhile, with RPIseq line computation lncRNA UCA1 and ovary cancer-associated protein (p53, ER, FGFR-1 and
BRCA1 etc.) between when potentially interacting, find the associated score between lncRNA UCA1 and each ovary cancer-associated protein
Above 0.5, and SVM prediction score is 0.95 or so, showing between lncRNA UCA1 and ovary cancer-associated protein that pole has can
There can be interaction, and the expression by adjusting ovary cancer-associated protein influences the progress of oophoroma, therefore lncRNA
UCA1 can as the biomarker of diagnosis of ovarian cancer, assist doctor to patient whether suffer from oophoroma or stages of ovarian carcinoma into
Row judgement；It can be used as the biomarker that outcome is examined after treatment of ovarian cancer simultaneously, auxiliary doctor is to taking treatment
The progress of oophoroma after means examines associated treatment means to ovarian cancer prognosis effect, so that the treatment for oophoroma provides
New approaches and new way；LncRNA UCA1 can be provided accurately in conjunction with other clinical indices for diagnosis, the prognosis of oophoroma
Believable inspection result.
(2) PCR primer and PCR kit provided by the present application for detecting lncRNA UCA1 expression has spirit
The features such as sensitivity is high, at low cost, detection process is simple.(3) PCR kit of the application is by extracting the total of patient tissue samples
Total serum IgE reverse transcription is cDNA by RNA, recycles the expression water of simple DNA dye method detection ovarian carcinoma associated genes UCA1
It is flat, various clinicopathological parameters are combined by the difference degree of the expression, the risk having ovarian cancer can be better understood by
Or the effect of prognosis.Therefore the kit puts into production practice, to distinguishing oophoroma and clinical auxiliary detection with important work
With, if oophoroma can be diagnosed to be in time, patient just have early stage cure chance, clinically for be also revolution
Breakthrough.
Detailed description of the invention
Fig. 1 is the result that in TCGA database 606 oophoroma cases are carried out with differential expression lncRNA screening；
Wherein, Genetic Alteration indicates " genetic alteration ", refers to the lncRNA with science of heredity variation；
Amplification indicates " gene magnification ", refers to that unconventionality expression, mRNA occurs in corresponding lncRNA in the tissue samples
Upregulaion indicates " mRNA expression up-regulation ", refers to that unconventionality expression, No alterations occurs in total mRNA in the tissue samples
It indicates " unchanged ", refers to that unconventionality expression does not occur in corresponding lncRNA in the tissue samples, Not profiled indicates no survey
Examination；" * % " indicates that the ratio of the total sample of tissue samples Zhan of unconventionality expression occurs in corresponding lncRNA；
Fig. 2 is the result predicted the interaction of lncRNA UCA1 and ovary cancer-associated protein；
Wherein, score indicates " score ", and RF indicates that Random forest classifier algorithm, SVM indicate
Support Vector Machine algorithm；
Fig. 3 is the lncRNA UCA1 that detects in conjunction with the GEO database chip sequencing result group by ovarian cancer tissue and cancer
The expression knitted；
Wherein, UCA1expression value (log2RMA signal intensity) indicates that lncRNA UCA1 exists
The fold differences of cancerous tissue and normal tissue expression, Normal indicate normal tissue, and Cancer indicates ovarian cancer tissue, P <
0.01 indicates lncRNA UCA1, and there are significant differential expressions in ovarian cancer tissue and normal tissue.
Specific embodiment
Further details of the technical solution of the present invention with reference to the accompanying drawings and detailed description.
The screening and determination of lncRNA biomarker in 1 oophoroma of embodiment
Screening target lncRNA is carried out using TCGA database, HGNC database, GEO database and qRT-PCR technology:
(1) lncRNA of all approved symbol is downloaded in HGNC database, totally 2773；
(2) all lncRNA are screened in TCGA database website, selects the gene expression atlas of ovarian neoplasm
MRNA Expression z-Scores (RNA Seq V2RSEM), the result of generation is ranked up, and wherein ovary is swollen for selection
Tumor science of heredity has several lncRNAs of change rate, is respectively: UCA1 (its nucleotide sequence is as shown in SEQ ID No.1),
FBXO43 and LINC01192 (as shown in Figure 1).
(3) result of TCGA database preliminary screening is compared with GEO database oophoroma chip sequencing result, is gone
Fall it is some without apparent differential expression as a result, final determining target lncRNA UCA1.
(3) ovarian cancer tissue and cancer beside organism are acquired within the hospital, extract its total serum IgE using RNA kit, and use
Nanodrop 2000 detects the content and quality of RNA, is uniformly diluted to 100ng/ul, volume needed for calculating reverse transcription.
(4) by the RNA reverse transcription of extraction be cDNA, reverse transcription reaction system such as table 1:
1 reverse transcription reaction system of table
(5) RT-q PCR is carried out in new 200 μ L optics PCR, eight union, each sample carries out three multiple holes and repeats,
RT-q PCR system such as table 2:
2 RT-q PCR system of table
By RT-q PCR system mix after the of short duration centrifugation several seconds, then be placed in ABI Prism 7500 carry out RT-qPCR it is anti-
It answers, reaction condition such as table 3:
3 RT-qPCR response procedures of table
(6) processing and analysis of data
When relative expression's variable quantity of quantitative fluorescent PCR quantitative detection lncRNA, using GAPDH as reference gene, to mesh
Mark lncRNA is normalized, to ensure to compare the expression quantity of target gene in the sample of equal quantity.Expression quantity
The formula of the variation of multiple are as follows:
RQ=2-△△Ct, △ △ Ct=(CtlncRNA-CtGAPDH)BC-(CtlncRNA-CtGAPDH) BA,
Wherein RQ represents relative expression quantity (Relative Quantitation), CtlncRNAAnd CtGAPDHRespectively represent fluorescence
The Ct value of target lncRNA and reference gene GAPDH that quantitative detection arrives, BC represent ovarian cancer tissue (Ovarian Cancer
Tissue), BA represents cancer beside organism corresponding with tumor tissues sample (Ovarian cancer Adjacent tissue).
Setting three repeated experiments and negative control experiment in quantitative: each sample is repeated 3 times in quantitative sample, negative right
It is added without sample form cDNA according in, but is replaced with water, for detecting whether there are PCR pollution or higher primer dimerization
Body pollution.
Statistical analysis is carried out to the data that quantitative fluorescent PCR obtains using SPSS17.0 statistical analysis software,
Relative expression quantity analysis of the lncRNA UCA1 in two tissue samples (ovarian cancer tissue and cancer beside organism) uses levene ' s
Chi-square Test and independent-sample t are examined, as P value < 0.05, it is believed that result statistically has conspicuousness
Difference.As P value < 0.01, it is believed that result statistically has extremely significant sex differernce (as shown in Figure 2).
Experimental data shows that there are significant differences for lncRNA UCA1 expression in ovarian cancer tissue and cancer beside organism, can be first
Step is used as oophoroma biomarker.
Embodiment 2
With RPIseq line computation lncRNA UCA1 protein relevant to oophoroma (p53, ER, FGFR-1 and
BRCA1 albumen) potentially interaction.
Two different calculation methods are used in the algorithm that RPIseq calculates interaction, one is RPIseq-
SVM, i.e. Support Vector Machine；Another kind is RPIseq-RF, i.e. Random forest classifier.Benefit
The result obtained with both algorithms the score (as shown in Figure 3) that can be seen that lncRNA UCA1 and each cancer-associated proteins
Above 0.5, and SVM prediction score, 0.95 or so, it is very big that this illustrates that UCA1 and p53, ER, FGFR-1 and BRCA1 albumen have
There may be interactions, and the progress of oophoroma may be influenced by adjusting the expression of ovary cancer-associated protein.
Embodiment 3 is used to detect the real-time fluorescence quantitative PCR kit of lncRNA UCA1 expression
It is a kind of for detecting the real-time fluorescence quantitative PCR kit of lncRNA UCA1 expression, comprising:
(1) reverse transcription system: M-MLV (H-) reverse transcriptase (200U/ μ L) and reverse transcription 5 × RT of system buffer buffering
Liquid, dNTP mixture (10nm each), gene specific Primers (2 μM), RNase inhibitor (40U/ μ L)；Wherein,
The nucleotide sequence of gene specific Primers (gene-specific primer) is as shown in SEQ ID No.3.
(2) PCR amplification system: qPCR reaction mixture (High ROX), UCA1 upstream amplification primer (such as SEQ ID
No.2), UCA1 downstream amplification primer (such as SEQ ID No.3), GAPDH upstream amplification primer (such as SEQ ID No.4), GAPDH
Downstream amplification primer (such as SEQ ID No.5).
(3)RNase free ddH2O。
Wherein, the RNA reverse transcription that reverse transcription system is used to extract is cDNA, and reverse transcription reaction system is the same as table 1.And PCR
The reaction system of amplification system such as table 4:
Table 4 is used to detect the PCR amplification system of the real-time fluorescence quantitative PCR kit of lncRNA UCA1 expression
Brief centrifugation after each reagent of PCR amplification system is mixed, is operated the computer, response parameter setting are as follows:
Stage is 1.: 95 DEG C, 5min；
Stage is 2.: 40 circular responses: 95 DEG C, 10s, 60 DEG C, 30s；
Stage is 3.: 95 DEG C, 15s, 60 DEG C, 1min, 95 DEG C, 15s.
SEQUENCE LISTING
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<120>lncRNA UCA1 is as the application in ovarian cancer diagnosis or the biomarker of outcome inspection
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aggcuucauc cguuccucug gacccucauc ucuuaagacc ugccgccuau aaaaaggauu 900
auaucuugag acccuauccu cuaaaauuuu uuccacaccc aaaacaaaaa aucucugggu 960
caaaagucua aaacgcuuag gcuggcaacc aucagauccu ugcccauggu guccucaagc 1020
cuacucucau gaaauggaca acaguacacg cauauggggc caguuccaca uauuuggcaa 1080
ccagaccagc auccaggaca acacaaagua uguuguuugu uguuagaggg cuugggacau 1140
uucacucuuu gccagccuca gcuuaaucca ggagacaaag auuauuuucc uuauuaucuc 1200
uucugcauag gaucugcaau cagaacuauu gaacuucucc auucagaccg ccacucacac 1260
cuaugggaaa aggguaaugu aucaucggcu uagcaacagg gaauacuauu cguaugaugg 1320
aaaaugggga caaaaggcuu ugguacauaa aacauuauuc cuuccuuggc cuaaaaacuc 1380
aucgccaccu acauuaaagc uaauaugccu gauuacuguu uuuagagaac uuauuuuauu 1440
agggcaguuc caagcucaaa aauacgcuaa cuggcaccuu guuagcuaca uaaaaaugca 1500
cccuagaccc gaaacuuacu agacucauua uaaaauuuuc uuuaaggugu ccacgcaguc 1560
ccuggucaca cuugaagcag uccggagaaa uaucagcccu accccaguaa uccccagaag 1620
gaacuuacac uuuuuuuuaa ucuuuuccua caacuucaua uuuuauaaau aaaaagacaa 1680
aaaugucagg ccugugagcu gaagcuuagc cauuguaacc ccugugaccu gcacauaucc 1740
guccaggugg ccugcaggag ccaagaaguc uggagcagcc gaaaaaccac aaagaaguga 1800
aacagccagu uccugccuua acuaauuaac ccaccuuacg acauuccacc auuaugacuu 1860
guccaccauu augacuuguu ccugcccugc cccaacugau caaucaaccc ugugacauuc 1920
uucuccugga caaugagucc caucaucucu ccaccaugca ccuugugacc cccuccucug 1980
cugaggauaa ccaccuuuaa cuguaacuuu ccacgccuac ccaagcccua uaaagcugcc 2040
ccucuccuau cucccuucac ugacucucuu uucggacuca gcccacuugc acccaaguga 2100
auuaacagcc uuguugcuca cacaaagccu gauugggugu cuucuauacg gacacgcgug 2160
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aggcuuuugu aaacagaggc guuucaugug guuuuccuuu ccuuuccuua uaugugaaaa 2280
ggugacagaa aagaaaucuu ccuaaaagag ucag 2314
<210> 2
<211> 29
<212> DNA
<213>artificial synthesized sequence
<400> 2
tcaatcaacc ctgtgacatt cttctcctg 29
<210> 3
<211> 33
<212> DNA
<213>artificial synthesized sequence
<400> 3
ggaagatttc ttttctgtca ccttttcaca tat 33
<210> 4
<211> 19
<212> DNA
<213>artificial synthesized sequence
<400> 4
aacggatttg gtcgtattg 19
<210> 5
<211> 19
<212> DNA
<213>artificial synthesized sequence
<400> 5
ggaagatggt gatgggatt 19
Claims (10)
1. a kind of application of lncRNA UCA1 in the biomarker as ovarian cancer diagnosis, which is characterized in that described
The nucleotide sequence of lncRNA UCA1 is as shown in SEQ ID No.1.
2. a kind of application of lncRNA UCA1 in the biomarker as ovarian cancer diagnosis, which is characterized in that described
Nucleotide sequence shown in the nucleotide sequence and SEQ ID No.1 of lncRNA UCA1 has 90% or more homology.
3. a kind of application of lncRNA UCA1 in the biomarker as ovarian cancer prognosis validity check, which is characterized in that
The nucleotide sequence of the lncRNA UCA1 is as shown in SEQ ID No.1.
4. a kind of application of lncRNA UCA1 in the biomarker as ovarian cancer prognosis validity check, which is characterized in that
The nucleotide sequence of the lncRNA UCA1 and nucleotide sequence shown in SEQ ID No.1 with 90% or more it is homologous
Property.
5. a kind of for detecting the PCR primer of lncRNA UCA1 expression characterized by comprising
UCA1 upstream primer: 5 '-TCAATCAACCCTGTGACATTCTTCTCCTG-3 '；
UCA1 downstream primer: 5 '-GGAAGATTTCTTTTCTGTCACCTTTTCACATAT-3 '.
6. a kind of for detecting the PCR kit of lncRNA UCA1 expression, which is characterized in that containing such as claim 5 institute
That states is used to detect the PCR primer of lncRNA UCA1 expression, and the PCR for detecting reference gene expression draws
Object.
7. as claimed in claim 6 for detecting the PCR kit of lncRNA UCA1 expression, which is characterized in that described
Reference gene be GAPDH gene, the described PCR primer for detecting reference gene expression includes:
GAPDH upstream primer: 5 '-AACGGATTTGGTCGTATTG-3 '；
GAPDH downstream primer: 5 '-GGAAGATGGTGATGGGATT-3 '.
8. as claimed in claim 7 for detecting the PCR kit of lncRNA UCA1 expression, which is characterized in that
The real-time fluorescence quantitative PCR reaction system of lncRNA UCA1 includes: the qPCR reaction mixture of 5 μ L, the upstream UCA1 of 0.5 μ L
Primer, the UCA1 downstream primer of 0.5 μ L, the GAPDH upstream primer of 0.5 μ L, the GAPDH downstream primer of 0.5 μ L, 1 μ L cDNA,
RNase free ddH2O complements to 10 μ L.
9. as claimed in claim 8 for detecting the PCR kit of lncRNA UCA1 expression, which is characterized in that
The real-time fluorescence quantitative PCR response procedures of lncRNA UCA1 are as follows:
Stage is 1.: 95 DEG C, 5min；
Stage is 2.: 40 circular responses: 95 DEG C, 10s, 60 DEG C, 30s；
Stage is 3.: 95 DEG C, 15s, 60 DEG C, 1min, 95 DEG C, 15s.
10. as claimed in claim 6 for detecting the PCR kit of lncRNA UCA1 expression, which is characterized in that also
The reverse transcription system for being cDNA containing the total serum IgE reverse transcription for that will extract, the reverse transcription system include M-MLV (H-) anti-
Transcriptase, 5 × RT M-MLV buffer, dNTP mixture, the UCA1 downstream primer and concentration that concentration is 2 μM are 40U/ μ L
RNase inhibitor；
Reverse transcription reaction system includes: 5 × RT M-MLV buffering of the total serum IgE of 2.5 μ L, the UCA1 downstream primer of 0.5 μ L, 3 μ L
Liquid, the dNTP mixture of 0.25 μ L, M-MLV (H-) reverse transcriptase of 0.5 μ L, 0.25 μ L RNase inhibitor,
Described M-MLV (H-) reverse transcriptase uses after being configured to the enzyme solutions that concentration is 200U/ μ L.
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