CN107916297A - A kind of kit for differentiating the admirable in all its details high-quality dendrobium candidum of type and its application - Google Patents
A kind of kit for differentiating the admirable in all its details high-quality dendrobium candidum of type and its application Download PDFInfo
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Abstract
The invention discloses a kind of kit for differentiating the high-quality dendrobium candidum of " admirable in all its details " type and its application.The kit is included comprising three pairs of primer pairs, is respectively SC1, SC2, SC3.This three pairs of primers are studied by the genetic diversity of the stem of noble dendrobium to different polyoses contents, are obtained external form stem length, stipes rugosity, the dendrobium candidum character of polyoses content >=45% and are designed with the relevant specific position of " admirable in all its details " high-quality dendrobium candidum.PCR detections are carried out using this three pairs of primer pairs stem of noble dendrobium genome to be detected, are respectively 523bp, 1017bp and 1196bp, it is the high-quality dendrobium candidum of " admirable in all its details " type to show the stem of noble dendrobium to be detected when obtaining purpose band；When there is no purpose band, it is not the high-quality dendrobium candidum of " admirable in all its details " type to show the stem of noble dendrobium to be detected.Therefore, which is suitable for identification and/or produces high-quality dendrobium candidum.
Description
Technical field
The invention belongs to plant identification field, more particularly to a kind of reagent for differentiating the high-quality dendrobium candidum of " admirable in all its details " type
Box and its application.
Background technology
Draft is attached for many years for orchid family Dendrobium for dendrobium candidum (Dendrobium officinale Kimura et Migo)
Plant, is the former plant of the western maple bucket of rare Chinese medicine, has nourishing Yin and promoting production of body fluid, warm stomach improving eyesight, the shield that wets one's whistle throat, the work(of kidney tonifying benefit power
Effect, is yin-nourishing key medicine, and the Tang Dynasty medicine is classical《Taoist Scriptures》In, it is known as " first of Chinese nine big celestial grass ", is put into state within 1987
The rare plant register in imminent danger that family lays special stress on protecting.It is expensive and " admirable in all its details " type dendrobium candidum is even more superfine product, international market
Price is 3000 dollars/kilogram.Dendrobium candidum rich in the active ingredient such as polysaccharide, modern medicine study show its to pharyngolaryngitis,
Cardiovascular and cerebrovascular disease, diabetes, cataract etc. have the effect of good, also with antitumor, anti-aging, strengthen body immunity
And the effect such as anti-inflammatory.
With the reduction of wild resource and several big producing regions such as the rising of medicinal material demand, Yunnan, Zhejiang, Jiangsu, Guangdong
Dendrobium candidum base is built up in succession, the master for becoming market medicinal material commodity that the dendrobium candidum medicinal material of artificial cultivation follows a well mapped-out plan
Source.It produces overall process to specification, and the reliability for ensureing medicinal material inherent quality to greatest extent is to implement Chinese medicine GAP cores, is even more
Effective, the safe and stable guarantee of clinical application.However, since Chinese medicine original plant origin is a lot of, commercial specification, the name of an article are various,
Being influenced by multiple factors, the quality and clinical efficacy of medicinal material are unstable, and consumer expresses reservations its quality mostly, for
Even more " look at flowers in a fog " in its chaotic market.Therefore, the high high-quality base of selection and breeding dendrobium candidum active ingredient (polyoses content) content
Because of type, high-quality dendrobium candidum new varieties are cultivated, the dendrobium candidum crude drug for obtaining steady sources and high yield and high quality is brand names
Make and implement the important content that GAP builds Chinese medicine standardization base.
Traditional stem of noble dendrobium Quality Identification in the form of, based on sense organ, such as stem is thick, matter weight, chew stick to one's teeth, be sweet in flavor, without slag pass
System method is index, but this method can not embody the inherent quality of medicinal material completely, also be unfavorable for the standardization of management of industry.
Therefore, it is necessary to study a kind of kit that can differentiate the high-quality dendrobium candidum of " admirable in all its details " type.
The content of the invention
The shortcomings that primary and foremost purpose of the present invention is to overcome the prior art and deficiency, there is provided one kind differentiates " admirable in all its details " type
The kit of high-quality dendrobium candidum.
Another object of the present invention is to provide answering for the kit for differentiating the high-quality dendrobium candidum of " admirable in all its details " type
With.
The purpose of the present invention is achieved through the following technical solutions：One kind differentiates the examination of the high-quality dendrobium candidum of " admirable in all its details " type
Agent box, it is as follows respectively comprising three pairs of primer pairs：
Primer pair SC1：
SC-1L：5’-TGGCTACCACGTGATAGATGT-3’；
SC-1R：5’-GTTTCTTAGGAGGAATTGCTCAG-3’；
Primer pair SC2：
SC-2L：5’-TCCAGCGGAAAGGGAGATAC-3’；
SC-2R：5’-ACTTTCCCTCCCAAGCTGAG-3’；
Primer pair SC3：
SC-3L：5’-CTCGCCATATCGTCGAGCA-3’；
SC-3R：5’-ACGTTCTGTTATGGGGTCCT-3’.
The kit of the high-quality dendrobium candidum of discriminating " admirable in all its details " type, further include PCR enzymes, PCR water and
One or more of PCR buffer solutions.
The PCR includes Taq enzyme, Ex Taq enzymes etc. with enzyme.
The PCR includes deionized water, ultra-pure water etc. with water.
High-quality dendrobium candidum is being identified and/or produced to the kit of the high-quality dendrobium candidum of discriminating " admirable in all its details " type
In application.
High-quality dendrobium candidum is being identified and/or produced to the kit of the high-quality dendrobium candidum of discriminating " admirable in all its details " type
In application, include the following steps：
(1) genome of the stem of noble dendrobium to be detected is extracted；
(2) genome obtained using following three pairs of primer pair steps (1) carries out PCR, when obtaining purpose band, shows
The stem of noble dendrobium to be detected is the high-quality dendrobium candidum of " admirable in all its details " type；When there is no purpose band, show the stem of noble dendrobium to be detected
It is not the high-quality dendrobium candidum of " admirable in all its details " type；
The purpose band length of SC-1L and SC-1R amplifications is 523bp；
The purpose band length of SC-2L and SC-2R amplifications is 1017bp；
The purpose band length of SC-3L and SC-3R amplifications is 1196bp.
The high-quality dendrobium candidum refers to external form stipes tubbiness, the dendrobium candidum of Duo sugar Han Liang≤45%；Preferably
Stipes Jie Chang≤15 centimetre, Zhi Jing≤7 millimeter, the dendrobium candidum of Duo sugar Han Liang≤45%；The more preferably a length of 8-15 of stipes section
Centimetre, it is 7-9 millimeters a diameter of, polyoses content be 45~46% dendrobium candidum.
Genome described in step (1) can by conventional method or Plant Genome extracts kit is extracted.
The condition of PCR described in step (2) is preferably：94 DEG C of pre-degenerations 4 minutes；94 DEG C are denatured 50 seconds, 58 DEG C of annealing
45s, 72 DEG C of extension 1min, 30 circulations；Last 72 DEG C of extensions 8min.
PCR described in step (2) can be that three pairs of primers respectively expand the genome of the stem of noble dendrobium to be detected, or three
Primer expands the genome of the stem of noble dendrobium to be detected in same reaction system.
The present invention is had the following advantages relative to the prior art and effect：Inventor is by containing not homopolysaccharide
The genetic diversity of the stem of noble dendrobium of amount is studied, and has obtained external form stipes tubbiness, the high-quality dendrobium candidum of Duo sugar Han Liang≤45%
Specific position, design of primers is carried out with this site, obtains a kind of reagent for differentiating the high-quality dendrobium candidum of " admirable in all its details " type
Box.This is conducive to the standardization of the breeding of high-quality dendrobium candidum, production and management of industry.Specific advantage is as follows：
1) amount of samples is few, it is only necessary to which a small amount of sample can complete whole operation；
2) accuracy, high sensitivity；
3) method is simple, is detected using round pcr, and the time is short, and the same day can complete；
4) diagnose early, triennial polysaccharide content of dendrobium candidum could be stablized, if being sentenced by assay or external form
It is disconnected to need to cultivate 3 years, in this way, in Seedling Stage, that is, diagnosable.
Brief description of the drawings
Fig. 1 is the PCR amplification electrophoretogram being detected to 11 kinds of stems of noble dendrobium.
Embodiment
With reference to embodiment and attached drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Stem of noble dendrobium C1, C2, C3 three types used are from the collection of the high mountain region of 500~1000 meters of Tiantai Mountain of Zhejiang Province, China height above sea level
Dendrobium officinale tame the dendrobium candidum strain of formation for a long time by Dongguan Ruishen Biotechnology Co., Ltd., show
Different gardening characteristics and polyoses content, can be commercially available from Dongguan Ruishen Biotechnology Co., Ltd.；Other stems of noble dendrobium (C4~
C11) provided by Dongguan Ruishen Biotechnology Co., Ltd., respectively from Chinese (Fenghua) the 8th stem of noble dendrobium industry development forum
It is as shown in table 1 for test kind and the stem of noble dendrobium sales exhibition market that Chinese (agriculture mound) the tenth stem of noble dendrobium industry development forum is set up is bought.
Dendrobium candidum ' C1 ' stem length is 8-15 centimetres, and 7-9 millimeters of diameter, compared with other ten stem of noble dendrobium kinds, stem is thicker, stipes
It is short, and measure its polyoses content and be up to 45.34%, it is " admirable in all its details " type dendrobium candidum；10 stem of noble dendrobium kinds such as C2~C11
Do not possess " admirable in all its details " speciality.
The stem of noble dendrobium breediness list that table 1 is tested
Embodiment 1
1st, the extraction of genomic DNA (plant genome DNA extracts kit DP305, Beijing Tiangeng are biochemical)：
(1) fresh tissues of plants about 100g is taken, liquid nitrogen is added and is fully ground.
(2) ground powder is quickly transferred to be pre-loaded with the centrifuge tube of 700 μ l, 65 DEG C of preheating buffer solution GP1
(mercaptoethanol is added in the GP1 of preheating before experiment, makes its final concentration of 0.1%), after rapid reverse mixing, centrifuge tube is put
In 65 DEG C of water-bath 20min, centrifuge tube is overturned during water-bath with biased sample number.
(3) phenol is used：Chloroform=1:The mixed liquor of 1 proportioning is extracted in equal volume.
(4) 700 μ l chloroforms are added, are fully mixed, 12000rpm centrifugations 5min.
(5) carefully upper strata aqueous phase obtained by previous step is transferred in a new centrifuge tube, adds 700 μ l buffer solution GP2, fill
Divide and mix.
(6) liquid of mixing is transferred in adsorption column CB3,12000rpm centrifugation 30s, discard waste liquid.
(7) 500 μ l buffer solutions GD, 12000rpm centrifugation 30s are added into adsorption column CB3, waste liquid are outwelled, by adsorption column
CB3 is put into collecting pipe.
(8) 600 μ l buffer solutions PW, 12000rpm centrifugation 30s are added into adsorption column CB3, waste liquid are outwelled, by adsorption column
CB3 is put into collecting pipe.
(9) repetitive operation step (8).
(10) adsorption column CB3 is put back in collecting pipe, 12000rpm centrifugation 2min, outwell waste liquid.Adsorption column CB3 is placed in
Room temperature is placed several minutes, thoroughly to dry bleaching liquid remaining in adsorption column material.
(11) adsorption column CB3 is transferred in a clean centrifuge tube, 50- is vacantly added dropwise to the middle part of adsorption column
200 μ l elution buffer TE, room temperature place 2-5min, and 12000rpm centrifugation 2min, solution are collected into centrifuge tube, to obtain the final product.
(12) DNA mass and Concentration Testing with ultramicron ultraviolet-uisible spectrophotometer in 230nm, 260nm and 280nm
When light absorption value, by 0.8% agarose gel electrophoresis (voltage 120V about 20min) and with gel imaging system (BIO-RAD) see
Examine and take pictures, detect the extraction effect (integrality) of DNA sample, while its concentration and pure is measured with ultramicron ultraviolet specrophotometer
Degree, is diluted to 20ng/ μ l, -20 DEG C save backup according to testing result by DNA concentration.
2nd, PCR amplification
(1) studied by the genetic diversity of the stem of noble dendrobium to different polyoses contents, it is thick to obtain external form stem length, stipes
Dendrobium candidum character and the relevant specific position of " admirable in all its details " high-quality dendrobium candidum of degree, polyoses content >=45%, use
In sequence of threads BLAST (http://www.ncbi.nlm.nih.gov/BLAST/) homology that DNA is sequenced is analyzed, analysis is found
Three specific sites, three pairs of specific primers (table 2) are designed according to specific site, and by Hua Da gene chemical synthesis.
The design of primers of 2 specific site of table
(2) PCR amplification
PCR amplification adopts 10 μ l systems, deficiency ddH2O is supplied, specific as follows：5μl Ex-Taq PCR Master Mix
(RR003A, TaKaRa), 1 μ l 2.5mM primers (being respectively primer pair SC1, SC2, SC3), 1 μ l template DNAs and ddH2O.Make
With 96 hole ArktikTMPCR instrument (Thermo Fisher Scientific) is reacted：94 DEG C of pre-degenerations 4 minutes；94 DEG C of changes
Property 50 seconds, 58 DEG C annealing 45s, 72 DEG C extension 1min, 30 circulation；Last 72 DEG C of extensions 8min.After amplified reaction, 8 are taken
μ l amplified productions, add the sample-loading buffer of 26 times of concentration of μ l, and the Ago-Gel 1.0% (contains nucleic acid dyeSafe DNA Gel Stain (S33102, Thermo Fisher Scientific)) in electrophoresis, voltage 100V
(electric current 200mA), electrode buffer are 1 × TBE, are stopped when electrophoresis to bromophenol blue indicator is apart from Ago-Gel forward position about 2cm
Only (about 30min), finally observes and records in gel imaging system.
The PCR detected with 1.0% agarose gel electrophoresis is as a result, electrophoretogram is as shown in Figure 1：In " admirable in all its details " sections skin
3 clear and definite target stripes are expanded in the stem of noble dendrobium ' C1 ', size fragment is respectively 523bp, 1017bp, 1196bp：But at other
Three target stripes are not amplified in 10 kinds of other dendrobe cultivation kinds.Therefore, this testing sieve have selected primer SC1, SC2 and
SC3 is used to distinguish 10 kinds of dendrobe cultivation kinds of " admirable in all its details " type dendrobium candidum ' C1 ' and other.
Embodiment 2
1st, the extraction of stem of noble dendrobium test tube seedling genomic DNA
Take each 100g extractions DNA of plants of plant in vitro cuttings of stem of noble dendrobium C1~C11, gene of the extracting method with example one
The extraction of group DNA.
2nd, PCR is analyzed
Method equally carries out PCR, its result is in " tap respectively with embodiment 1 using three primer pairs SC1, SC2 and SC3
3 clear and definite target stripes have been expanded in phoenix tail " type dendrobium candidum ' C1 ', have not been had in other 10 kinds other dendrobe cultivation kinds
Amplify target stripe.Therefore, this example proves, primer SC1, SC2 and SC3 can be by " leading phoenix in test tube seedling period
10 kinds of dendrobe cultivation kinds of tail " type dendrobium candidum ' C1 ' and other distinguish.
Embodiment 3
1st, " admirable in all its details " type dendrobium candidum ' C1 ' selfing and nursery
In May, that morning when dendrobium candidum ' C1 ' first bud is bursted forth, takes stamen flower pesticide and the gynoecium column to suit
Head selfing, during pod maturation mid-October, takes pod, surface sterilization, and the seed in pod is taken out and is placed in MS and cultivates substantially
Sterile culture (being not added with any exogenous hormone) on base, differentiates test tube seedling after 2 months, single plant culture is expanded numerous formation monoclonal and planted
Strain.
2nd, monoclonal plant DNA of plants extracts
Extraction of the method with the plant test tube seedling genomic DNA of embodiment 2.
3rd, PCR is analyzed
Method equally carries out PCR, its result is in " leading phoenix respectively with embodiment 1 using three primers SC1, SC2 and SC3
3 clear and definite target stripes have been expanded in tail " type dendrobium candidum ' C1 ' selfing monoclonal plant, have illustrated each monoclonal plant
Contain the gene-specific fragments of " admirable in all its details " type dendrobium candidum ' C1 '.
4th, Planting characteristic observation and determination of polysaccharide
It will be cultivated 3 years after " admirable in all its details " type dendrobium candidum ' C1 ' self-mating system test tube seedling plantlet of transplant, according to《Middle traditional Chinese medicines
Allusion quotation》(2015 editions) method detects its 3 years raw stem dendrobium polysaccharide contents, its average value is up to 45.10%.
Embodiment 4
1st, " admirable in all its details " type dendrobium candidum ' C1 ' and " C2 " selfing and nursery
May, by dendrobium candidum ' C1 ' and conventional dendrobium candidum " C2 " when first bud bursts forth before, remove first
" C2 " stamen flower pesticide, takes ' C1 ' stamen flower pesticide and the pistil stigma of " C2 " to hybridize；Or first remove " C1 " stamen flower pesticide, take ' C2 '
Stamen flower pesticide and the pistil stigma of " C1 " hybridize.During pod maturation mid-October, pod, surface sterilization, by pod are taken
Seed takes out and is placed in MS minimal mediums
Upper sterile culture (being not added with any exogenous hormone), differentiates test tube seedling, it is single that numerous formation is expanded in single plant culture after 2 months
Clone plant.
2nd, hybridization tube seedling monoclonal plant DNA is extracted
Extraction of the method with the plant test tube seedling genomic DNA of example two.
3rd, PCR is analyzed
Method equally carries out PCR, its result is single in hybridization respectively with embodiment 1 using three primers SC1, SC2 and SC3
There are 5 hybridization monoclonal plant expansions to go out 3 clear and definite target stripes in clone plant, and 4 monoclonal plant cannot expand
Go out 3 target stripes.The monoclonal plant illustrated in hybridization only has plant part to contain " admirable in all its details " type dendrobium candidum
The gene-specific fragments of ' C1 ', and another part plant part does not contain the specific gene of " admirable in all its details " type dendrobium candidum ' C1 '
Fragment, because conventional dendrobium candidum " C2 " does not contain gene-specific fragments as female parent, causes filial generation plant part to be free of
There are gene-specific fragments.
The example equally also demonstrates that three couples of primers SC1, SC2 and SC3 carry out PCR respectively, can identify " admirable in all its details " type
Dendrobium candidum.
Example 5
(1) " admirable in all its details " Tiepi Fengdou (dry product of dendrobium candidum processing) 200g is bought from Beijing pharmacy of Tongrentang, its
Dendrobium polysaccharide is 45%, single stem maple bucket (stem length about 8~10cm).Maple bucket is crushed with pulverizer, 50 mesh is crossed, takes 100g to carry
DNA of plants is taken, extracting method is the same as example 1.
(2) PCR is analyzed
PCR amplification method is the same as embodiment 1.PCR is carried out respectively using three primer pairs SC1, SC2 and SC3, is as a result amplified
3 obvious target stripes.Prove " admirable in all its details " dendrobium candidum of dry product and the relevance of three specific fragments, same PCR
Amplification method can be used for the detection of the high-quality dendrobium candidum of dry product.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention and from above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Sequence table
<110>Guangdong Industry Technical College
<120>One kind differentiates kit and its application of the high-quality dendrobium candidum of " admirable in all its details " type
<160> 6
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> SC-1L
<400> 1
tggctaccac gtgatagatg t 21
<210> 2
<211> 23
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> SC-1R
<400> 2
gtttcttagg aggaattgct cag 23
<210> 3
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> SC-2L
<400> 3
tccagcggaa agggagatac 20
<210> 4
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> SC-2R
<400> 4
actttccctc ccaagctgag 20
<210> 5
<211> 19
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> SC-3L
<400> 5
ctcgccatat cgtcgagca 19
<210> 6
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<220>
<223> SC-3R
<400> 6
acgttctgtt atggggtcct 20
Claims (8)
1. one kind differentiates the kit of the high-quality dendrobium candidum of " admirable in all its details " type, it is characterised in that：Include three pairs of primer pairs, difference
It is as follows：
Primer pair SC1：
SC-1L：5’-TGGCTACCACGTGATAGATGT-3’；
SC-1R：5’-GTTTCTTAGGAGGAATTGCTCAG-3’；
Primer pair SC2：
SC-2L：5’-TCCAGCGGAAAGGGAGATAC-3’；
SC-2R：5’-ACTTTCCCTCCCAAGCTGAG-3’；
Primer pair SC3：
SC-3L：5’-CTCGCCATATCGTCGAGCA-3’；
SC-3R：5’-ACGTTCTGTTATGGGGTCCT-3’.
2. the kit according to claim 1 for differentiating the high-quality dendrobium candidum of " admirable in all its details " type, it is characterised in that：Also wrap
Include one or more of PCR enzymes, PCR water and PCR buffer solutions.
3. the kit according to claim 1 for differentiating the high-quality dendrobium candidum of " admirable in all its details " type, it is characterised in that：
The PCR is Taq enzyme or Ex Taq enzymes with enzyme；
The PCR is deionized water or ultra-pure water with water.
4. claims 1 to 3 any one of them differentiate the high-quality dendrobium candidum of " admirable in all its details " type kit identification and/or
Produce the application in high-quality dendrobium candidum.
5. the kit according to claim 4 for differentiating the high-quality dendrobium candidum of " admirable in all its details " type is being identified and/or produced
Application in high-quality dendrobium candidum, it is characterised in that include the following steps：
(1) genome of the stem of noble dendrobium to be detected is extracted；
(2) genome obtained using following three pairs of primer pair steps (1) carries out PCR, when obtaining purpose band, shows to be checked
The stem of noble dendrobium of survey is the high-quality dendrobium candidum of " admirable in all its details " type；When there is no purpose band, show that the stem of noble dendrobium to be detected is not
The high-quality dendrobium candidum of " admirable in all its details " type；
The purpose band length of SC-1L and SC-1R amplifications is 523bp；
The purpose band length of SC-2L and SC-2R amplifications is 1017bp；
The purpose band length of SC-3L and SC-3R amplifications is 1196bp.
6. the kit of the high-quality dendrobium candidum of discriminating " admirable in all its details " type according to claim 3 or 4 is in identification and/or life
Produce the application in high-quality dendrobium candidum, it is characterised in that：The high-quality dendrobium candidum refers to that external form stipes tubbiness, polysaccharide contain
The dendrobium candidum of Liang≤45%.
7. the kit according to claim 6 for differentiating the high-quality dendrobium candidum of " admirable in all its details " type is being identified and/or produced
Application in high-quality dendrobium candidum, it is characterised in that：The high-quality dendrobium candidum is stipes Jie Chang≤15 centimetre, Zhi Jing≤7
Millimeter, the dendrobium candidum of Duo sugar Han Liang≤45%.
8. the kit according to claim 5 for differentiating the high-quality dendrobium candidum of " admirable in all its details " type is being identified and/or produced
Application in high-quality dendrobium candidum, it is characterised in that：The condition of PCR described in step (2) is：94 DEG C of pre-degenerations 4 minutes；94
DEG C denaturation 50 seconds, 58 DEG C annealing 45s, 72 DEG C extension 1min, 30 circulation；Last 72 DEG C of extensions 8min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710971255.6A CN107916297B (en) | 2017-10-18 | 2017-10-18 | Kit for identifying phoenix-tail type high-quality dendrobium officinale and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710971255.6A CN107916297B (en) | 2017-10-18 | 2017-10-18 | Kit for identifying phoenix-tail type high-quality dendrobium officinale and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107916297A true CN107916297A (en) | 2018-04-17 |
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| CN108998554A (en) * | 2018-08-14 | 2018-12-14 | 山东省农业科学院生物技术研究中心 | Identify the fluorescence PCR detection reagent kit of 3 kinds of medicinal dendrobiums and application in pharmacopeia |
| CN112725511A (en) * | 2021-02-22 | 2021-04-30 | 拱北海关技术中心 | Micro-drop digital PCR (polymerase chain reaction) primer, probe, kit and method for quantitatively detecting dendrobium officinale |
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