CN102154264B - Method for rapidly extracting total ribonucleic acid from blood - Google Patents
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Abstract
The invention relates to a method for rapidly extracting total ribonucleic acid from blood, belonging to the field of nucleic acid purification. The method comprises the following steps of: firstly, adding a rapid erythrocyte lysing solution into whole blood; performing centrifugal separation on a mixture of the rapid erythrocyte lysing solution and the whole blood; taking precipitate formed by the centrifugal separation out; adding suspension into the precipitate; adding a lysing solution and a protease K solution in sequence; uniformly mixing and then adding absolute ethyl alcohol; uniformly mixing and transferring into a silicon membrane adsorption column to perform adsorptive centrifugation; adding a deproteinizing solution and a deoxyribonuclease solution; performing deproteinizing centrifugation twice; adding a rinsing solution to perform desalting centrifugation; and finally performing drying centrifugation and adding water without ribonuclease, and eluting and centrifuging so as to obtain a ribonucleic acid solution. The method provided by the invention has the characteristics of high efficiency, rapidness and simplicity; and purified RNA (Ribonucleic Acid) can be used for various downstream experiments such as chip analysis, in vitro translation, molecular cloning and the like.
Description
Technical field
The present invention relates to the method for the total Yeast Nucleic Acid of a kind of rapid extraction blood, belong to field of nucleic acid purification.
Background technology
Peripheral blood is being brought into play important effect in immunoreation and the metabolism in the humans and animals body, because it is prone to acquiredly, in basis and clinical study, have very consequence as a kind of special organization.Peripheral blood can be used as molecular marker in the disease of hematopoietic system research and the exploitation that is used for extracorporeal diagnostic system, also can be used for the molecule monitoring of the outer numerous systemic diseases of blood system.Wherein, effectively extract a most important step in peripheral blood Yeast Nucleic Acid (hereinafter to be referred as the RNA) process.But because the easy solidifiability of blood itself; And the complicacy of the interior various kinds of cell of blood; With and unlike solid tissue that works extracts RNA again after frozen; Make that the extraction of the high-quality RNA of blood is very difficult, when especially the applying gene chip carries out the scanning of full gene expression profile to it, just more strict to quality and the quantitative requirement of RNA.
One of the biggest problem that extraction RNA faces from blood is that RNA is very unstable: research is illustrated in the promptly rapid and significantly degraded of RNA in the short several hrs after the blood sampling, and part RNA is induced after blood sampling in addition, and expressing increases.This RNA degraded and gene in vitro abduction delivering can cause when analyzing vivo gene transcript abundance, makeing mistakes, even low-abundance information dropout.And traditional centrifugal post method extraction blood rna needs from whole blood, isolate white corpuscle earlier, and needs could begin and the adsorption column combination behind the filtration step.With complex steps before adsorption column combines, and need more than 40 minutes.
Summary of the invention
The objective of the invention is to propose the method for the total Yeast Nucleic Acid of a kind of rapid extraction blood; Adopt a kind of new quick erythrocyte cracked liquid that red cell membrane cracking and blood plasma are disperseed; And can form the complex body deposition with RNA at once through cetyl trimethyl ammonium sulfate composition; With the integrity of assurance Yeast Nucleic Acid, and make leaching process efficient, quick.
The method of the total Yeast Nucleic Acid of rapid extraction blood that the present invention proposes may further comprise the steps:
(1) in 100～400 microlitre whole bloods, add quick erythrocyte cracked liquid, the volume ratio of adding is: whole blood: erythrocyte cracked liquid=1: 5, put upside down mixing 8～10 times, and obtain mixture;
(2) said mixture is carried out spinning, the rotating speed of spinning is 7300 rev/mins, and the spinning time is 3 minutes, takes out the throw out of spinning;
(3) in above-mentioned throw out, add 240 microlitre suspension-s, be ground to deposition and dissolve fully, add 200 microlitre lysates, mixing adds 20 microlitre Proteinase K solution again, fully puts upside down mixing, and 55 ℃ of held 10 minutes are put upside down mixing, obtain solution;
(4) in above-mentioned solution, add 240 microlitre absolute ethyl alcohols, change over to behind the mixing in the silicon fiml adsorption column, adsorb centrifugally, the rotating speed of centrifugal absorption is 12000 rev/mins, and centrifugal adsorption time is 1 minute, removes waste liquid;
(5) in the silicon fiml adsorption column of step (4), add 350 microlitre protein liquid removals, it is centrifugal to carry out first time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins, and the Deproteinization centrifugation time is 15 seconds, removes waste liquid;
(6) in the silicon fiml adsorption column of step (5), adding activity unit is dnase-solution 80 microlitres of 0.375Kunitz; After the room temperature held 15 minutes; Add 350 microlitre protein liquid removals again, it is centrifugal to carry out second time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins; The Deproteinization centrifugation time is 15 seconds, removes waste liquid;
(7) in the silicon fiml adsorption column of step (6), add 500 microlitre rinsing liquids, left standstill under the room temperature 2 minutes, it is centrifugal to carry out desalination, and the desalination centrifugal rotation speed is 12000 rev/mins, and the desalination centrifugation time is 15 seconds, removes waste liquid; Repeat this step once;
(8) it is centrifugal the silicon fiml adsorption column of step (7) to be carried out drying, and dry centrifugal rotation speed is 12000 rev/mins, and dry centrifugation time is 2 minutes, and adsorption column was placed under the room temperature 2～5 minutes, removes the rinsing liquid in the sorbing material;
(9) in the silicon fiml adsorption column of step (8), add the water of 30～50 microlitre deoxyribonucleases, the room temperature held is after 2 minutes, and it is centrifugal to carry out wash-out, and the wash-out centrifugal rotation speed is 12000 rev/mins, and the wash-out centrifugation time is 2 minutes, obtains rna solution.
In the aforesaid method, the quality of each component of described quick erythrocyte cracked liquid is:
Ammonium chloride 60～100 grams
Saleratus 15～20 grams
Tetrasodium ethylenediamine tetraacetate 6～8 grams
Cetyl trimethyl ammonium sulfate 5～10 grams
Above-mentioned each component back of weighing is mixed, add deionized water, obtain mixed solution, transfer pH value to 7.2～7.4 of mixed solution, and to make mixeding liquid volume be 1000 milliliters with sodium hydroxide solution.
In the aforesaid method, the prescription of described suspension-s is:
Sodium tartrate 11.5～17.25 grams
Guanidinium hydrochloride 573～764 grams
5～10 milliliters of Nonidet P40s (NP40)
Above-mentioned each component is measured the back mix, add deionized water, and to make mixeding liquid volume be 1000 milliliters
The method of the total Yeast Nucleic Acid of rapid extraction blood that the present invention proposes, its advantage is:
1, the inventive method has been used a kind of new quick erythrocyte cracked liquid; Therefore make the leaching process of the total Yeast Nucleic Acid of blood have efficient, quick, succinct characteristics; The direct total Yeast Nucleic Acid of separation and purification from whole blood; Need not to carry out earlier leukocytic separation, for example from 100 microliters of mouse whole bloods, extract total Yeast Nucleic Acid among the embodiment 2 of the inventive method and only need 25 minutes.
2, use total Yeast Nucleic Acid good in integrity of the inventive method purifying; Purity is high, can be used for multiple downstream experiments such as inverse transcription polymerase chain reaction (RT-PCR), real time fluorescent quantitative inverse transcription polymerase chain reaction (Real Time RT-PCR), chip analysis, the hybridization of the Yeast Nucleic Acid marking (Northern blot), dot blot (Dot blot), external translation, rnase protection analysis and molecular cloning.
Description of drawings
Fig. 1 is the embodiment 1 of the inventive method, from 400 microlitre people whole bloods, extracts the electrophoresis detection figure of total Yeast Nucleic Acid.
Wherein, swimming lane 1: dna molecules amount standard (MarkerIII).Swimming lane 2: the total Yeast Nucleic Acid that uses the inventive method purifying.
Fig. 2 is the embodiment 2 of the inventive method, from 100 microliters of mouse whole bloods, extracts the electrophoresis detection figure of total Yeast Nucleic Acid.
Wherein, swimming lane 1: dna molecules amount standard (MarkerIII).Swimming lane 2: the total Yeast Nucleic Acid that uses the inventive method purifying.
Embodiment
The method of the total Yeast Nucleic Acid of rapid extraction blood that the present invention proposes may further comprise the steps:
(1) in 100～400 microlitre whole bloods, add quick erythrocyte cracked liquid, the volume ratio of adding is: whole blood: erythrocyte cracked liquid=1: 5, put upside down mixing 8～10 times, and obtain mixture;
(2) said mixture is carried out spinning, the rotating speed of spinning is 7300 rev/mins, and the spinning time is 3 minutes, takes out the throw out of spinning;
(3) in above-mentioned throw out, add 240 microlitre suspension-s, be ground to deposition and dissolve fully, add 200 microlitre lysates, mixing adds 20 microlitre Proteinase K solution again, fully puts upside down mixing, and 55 ℃ of held 10 minutes are put upside down mixing, obtain solution;
(4) in above-mentioned solution, add 240 microlitre absolute ethyl alcohols, change over to behind the mixing in the silicon fiml adsorption column, adsorb centrifugally, the rotating speed of centrifugal absorption is 12000 rev/mins, and centrifugal adsorption time is 1 minute, removes waste liquid;
(5) in the silicon fiml adsorption column of step (4), add 350 microlitre protein liquid removals, it is centrifugal to carry out first time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins, and the Deproteinization centrifugation time is 15 seconds, removes waste liquid;
(6) in the silicon fiml adsorption column of step (5), adding activity unit is dnase-solution 80 microlitres of 0.375Kunitz; After the room temperature held 15 minutes; Add 350 microlitre protein liquid removals again, it is centrifugal to carry out second time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins; The Deproteinization centrifugation time is 15 seconds, removes waste liquid;
(7) in the silicon fiml adsorption column of step (6), add 500 microlitre rinsing liquids, left standstill under the room temperature 2 minutes, it is centrifugal to carry out desalination, and the desalination centrifugal rotation speed is 12000 rev/mins, and the desalination centrifugation time is 15 seconds, removes waste liquid; Repeat this step once;
(8) it is centrifugal the silicon fiml adsorption column of step (7) to be carried out drying, and dry centrifugal rotation speed is 12000 rev/mins, and dry centrifugation time is 2 minutes, and adsorption column was placed under the room temperature 2～5 minutes, removes the rinsing liquid in the sorbing material;
(9) in the silicon fiml adsorption column of step (8), add the water of 30～50 microlitre deoxyribonucleases, the room temperature held is after 2 minutes, and it is centrifugal to carry out wash-out, and the wash-out centrifugal rotation speed is 12000 rev/mins, and the wash-out centrifugation time is 2 minutes, obtains rna solution.
In the aforesaid method, the quality of each component of described quick erythrocyte cracked liquid is:
Ammonium chloride 60～100 grams
Saleratus 15～20 grams
Tetrasodium ethylenediamine tetraacetate 6～8 grams
Cetyl trimethyl ammonium sulfate 5～10 grams
Above-mentioned each component back of weighing is mixed, add deionized water, obtain mixed solution, transfer pH value to 7.2～7.4 of mixed solution, and to make mixeding liquid volume be 1000 milliliters with sodium hydroxide solution.
In the aforesaid method, the prescription of described suspension-s is:
Sodium tartrate 11.5～17.25 grams
Guanidinium hydrochloride 573～764 grams
5～10 milliliters of Nonidet P40s (NP40)
Above-mentioned each component is measured the back mix, add deionized water, and to make mixeding liquid volume be 1000 milliliters.
Below introduce the embodiment of the inventive method.
Embodiment 1: from people's whole blood, extract total RNA.
(1) in 400 microlitre people whole bloods, add quick erythrocyte cracked liquid, the volume ratio of adding is: whole blood: erythrocyte cracked liquid=1: 5, put upside down mixing 8～10 times, and obtain mixture.Erythrocyte cracked liquid wherein; Its preparation method is: ammonium chloride 65 grams, saleratus 18 grams, tetrasodium ethylenediamine tetraacetate 6.5g and cetyl trimethyl ammonium sulfate 9 grams are mixed; Add deionized water; Obtain mixed solution, transfer the pH value to 7.3 of mixed solution, and to make mixeding liquid volume be 1000 milliliters with sodium hydroxide solution.
(2) said mixture is carried out spinning, the rotating speed of spinning is 7300 rev/mins, and the spinning time is 10 minutes, takes out the throw out of spinning.
(3) in above-mentioned throw out, add 10 milliliters of 240 microlitre suspension-s above-mentioned each component is measured the back mixing, add deionized water, and to make mixeding liquid volume be 1000 milliliters); Be ground to deposition and dissolve fully, add 200 microlitre lysates, mixing; Add 20 microlitre Proteinase K solution again, fully put upside down mixing, 55 ℃ of held 10 minutes; Put upside down mixing, obtain solution.Suspension-s wherein, its preparation method is: with sodium tartrate 12 gram, Guanidinium hydrochloride 573 grams and 10 milliliters of mixing of Nonidet P40 (NP40), add deionized water, and to make mixeding liquid volume be 1000 milliliters.
(4) in above-mentioned solution, add 240 microlitre absolute ethyl alcohols, change over to behind the mixing in the silicon fiml adsorption column, adsorb centrifugally, the rotating speed of centrifugal absorption is 12000 rev/mins, and centrifugal adsorption time is 1 minute, removes waste liquid.
(5) in the silicon fiml adsorption column of step (4), add 350 microlitre protein liquid removals, it is centrifugal to carry out first time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins, and the Deproteinization centrifugation time is 15 seconds, removes waste liquid.
(6) in the silicon fiml adsorption column of step (5), adding activity unit is dnase-solution 80 microlitres of 0.375Kunitz; After the room temperature held 15 minutes; Add 350 microlitre protein liquid removals again, it is centrifugal to carry out second time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins; The Deproteinization centrifugation time is 15 seconds, removes waste liquid.
(7) in the silicon fiml adsorption column of step (6), add 500 microlitre rinsing liquids, left standstill under the room temperature 2 minutes, it is centrifugal to carry out desalination, and the desalination centrifugal rotation speed is 12000 rev/mins, and the desalination centrifugation time is 15 seconds, removes waste liquid; Repeat this step once.
(8) it is centrifugal the silicon fiml adsorption column of step (7) to be carried out drying, and dry centrifugal rotation speed is 12000 rev/mins, and dry centrifugation time is 2 minutes, and adsorption column was placed under the room temperature 2～5 minutes, removes the rinsing liquid in the sorbing material;
(9) in the silicon fiml adsorption column of step (8), add the water of 30～50 microlitre deoxyribonucleases, the room temperature held is after 2 minutes, and it is centrifugal to carry out wash-out, and the wash-out centrifugal rotation speed is 12000 rev/mins, and the wash-out centrifugation time is 2 minutes, obtains rna solution.
Shown in Figure 1 is the electrophoresis detection figure that from 400 microlitre people whole bloods, extracts total Yeast Nucleic Acid among the embodiment 1, and swimming lane 1 among the figure: dna molecules amount standard (MarkerIII), swimming lane 2: the total Yeast Nucleic Acid that uses the inventive method purifying.
Embodiment 2: from the whole blood of mouse, extract total RNA.
(1) in 100 microliters of mouse whole bloods, add quick erythrocyte cracked liquid, the volume ratio of adding is: whole blood: erythrocyte cracked liquid=1: 5, put upside down mixing 8～10 times, and obtain mixture.Erythrocyte cracked liquid wherein; Its preparation method is: ammonium chloride 65 grams, saleratus 18 grams, tetrasodium ethylenediamine tetraacetate 6.5g and cetyl trimethyl ammonium sulfate 9 grams are mixed; Add deionized water; Obtain mixed solution, transfer the pH value to 7.3 of mixed solution, and to make mixeding liquid volume be 1000 milliliters with sodium hydroxide solution.
(2) said mixture is carried out spinning, the rotating speed of spinning is 7300 rev/mins, and the spinning time is 3 minutes, takes out the throw out of spinning;
(3) in above-mentioned throw out, add 240 microlitre suspension-s, be ground to deposition and dissolve fully, add 200 microlitre lysates, mixing adds 20 microlitre Proteinase K solution again, fully puts upside down mixing, and 55 ℃ of held 10 minutes are put upside down mixing, obtain solution.Wherein the preparation method of suspension-s is: sodium tartrate 12 gram, Guanidinium hydrochloride 573 grams and 10 milliliters of mixing of Nonidet P40 (NP40) add deionized water, and to make mixeding liquid volume are 1000 milliliters.
(4) in above-mentioned solution, add 240 microlitre absolute ethyl alcohols, change over to behind the mixing in the silicon fiml adsorption column, adsorb centrifugally, the rotating speed of centrifugal absorption is 12000 rev/mins, and centrifugal adsorption time is 1 minute, removes waste liquid.
(5) in the silicon fiml adsorption column of step (4), add 350 microlitre protein liquid removals, it is centrifugal to carry out first time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins, and the Deproteinization centrifugation time is 15 seconds, removes waste liquid.
(6) in the silicon fiml adsorption column of step (5), adding activity unit is dnase-solution 80 microlitres of 0.375Kunitz; After the room temperature held 15 minutes; Add 350 microlitre protein liquid removals again, it is centrifugal to carry out second time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins; The Deproteinization centrifugation time is 15 seconds, removes waste liquid.
(7) in the silicon fiml adsorption column of step (6), add 500 microlitre rinsing liquids, left standstill under the room temperature 2 minutes, it is centrifugal to carry out desalination, and the desalination centrifugal rotation speed is 12000 rev/mins, and the desalination centrifugation time is 15 seconds, removes waste liquid; Repeat this step once.
(8) it is centrifugal the silicon fiml adsorption column of step (7) to be carried out drying, and dry centrifugal rotation speed is 12000 rev/mins, and dry centrifugation time is 2 minutes, and adsorption column was placed under the room temperature 2～5 minutes, removes the rinsing liquid in the sorbing material;
(9) in the silicon fiml adsorption column of step (8), add the water of 30～50 microlitre deoxyribonucleases, the room temperature held is after 2 minutes, and it is centrifugal to carry out wash-out, and the wash-out centrifugal rotation speed is 12000 rev/mins, and the wash-out centrifugation time is 2 minutes, obtains rna solution.
Shown in Figure 2 is the electrophoresis detection figure that from 100 microliters of mouse whole bloods, extracts total Yeast Nucleic Acid among the embodiment 2, and wherein swimming lane 1: dna molecules amount standard (MarkerIII), swimming lane 2: the total Yeast Nucleic Acid that uses the inventive method purifying.
In the foregoing description of the inventive method, used lysate, silicon fiml adsorption column, dnase-solution, protein liquid removal, rinsing liquid and dna molecular amount standard (Marker III) etc. are produced by TIANGEN Biotech (Beijing) Co., Ltd..
Claims (1)
1. the method for the total Yeast Nucleic Acid of rapid extraction blood is characterized in that this method comprises following each step:
(1) in 100～400 microlitre whole bloods, add quick erythrocyte cracked liquid, the volume ratio of adding is: whole blood: erythrocyte cracked liquid=1: 5, put upside down mixing 8～10 times, and obtain mixture, the quality of each component of wherein said quick erythrocyte cracked liquid is:
Ammonium chloride 60～100 grams
Saleratus 15～20 grams
Tetrasodium ethylenediamine tetraacetate 6～8 grams
Cetyl trimethyl ammonium sulfate 5～10 grams
Above-mentioned each component back of weighing is mixed, add deionized water, obtain mixed solution, transfer pH value to 7.2～7.4 of mixed solution, and to make mixeding liquid volume be 1000 milliliters with sodium hydroxide solution;
(2) said mixture is carried out spinning, the rotating speed of spinning is 7300 rev/mins, and the spinning time is 3 minutes, takes out the throw out of spinning;
(3) in above-mentioned throw out, add 240 microlitre suspension-s, be ground to deposition and dissolve fully, add 200 microlitre lysates, mixing; Add 20 microlitre Proteinase K solution again, fully put upside down mixing, 55 ℃ of held 10 minutes; Put upside down mixing, obtain solution, the prescription of wherein said suspension-s is:
Sodium tartrate 11.5～17.25 grams
Guanidinium hydrochloride 573～764 grams
5～10 milliliters of Nonidet P40s
Above-mentioned each component is measured the back mix, add deionized water, and to make mixeding liquid volume be 1000 milliliters;
(4) in above-mentioned solution, add 240 microlitre absolute ethyl alcohols, change over to behind the mixing in the silicon fiml adsorption column, adsorb centrifugally, the rotating speed of centrifugal absorption is 12000 rev/mins, and centrifugal adsorption time is 1 minute, removes waste liquid;
(5) in the silicon fiml adsorption column of step (4), add 350 microlitre protein liquid removals, it is centrifugal to carry out first time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins, and the Deproteinization centrifugation time is 15 seconds, removes waste liquid;
(6) in the silicon fiml adsorption column of step (5), adding activity unit is dnase-solution 80 microlitres of 0.375Kunitz; After the room temperature held 15 minutes; Add 350 microlitre protein liquid removals again, it is centrifugal to carry out second time Deproteinization, and the Deproteinization centrifugal rotational speed is 12000 rev/mins; The Deproteinization centrifugation time is 15 seconds, removes waste liquid;
(7) in the silicon fiml adsorption column of step (6), add 500 microlitre rinsing liquids, left standstill under the room temperature 2 minutes, it is centrifugal to carry out desalination, and the desalination centrifugal rotation speed is 12000 rev/mins, and the desalination centrifugation time is 15 seconds, removes waste liquid; Repeat this step once;
(8) it is centrifugal the silicon fiml adsorption column of step (7) to be carried out drying, and dry centrifugal rotation speed is 12000 rev/mins, and dry centrifugation time is 2 minutes, and adsorption column was placed under the room temperature 2～5 minutes, removes the rinsing liquid in the sorbing material;
(9) in the silicon fiml adsorption column of step (8), add the water of 30～50 microlitre deoxyribonucleases, the room temperature held is after 2 minutes, and it is centrifugal to carry out wash-out, and the wash-out centrifugal rotation speed is 12000 rev/mins, and the wash-out centrifugation time is 2 minutes, obtains rna solution.
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| JP2014526255A (en) * | 2011-09-13 | 2014-10-06 | キアゲン ゲーエムベーハー | Method for isolating nucleic acids from veterinary whole blood samples |
| CN102586231B (en) * | 2012-03-07 | 2013-12-18 | 天根生化科技（北京）有限公司 | Rapid large-segment genome desoxyribonucleic acid extraction method |
| CN102888397A (en) * | 2012-09-25 | 2013-01-23 | 杭州硕航生物科技有限公司 | Kit using magnetic bead to extract whole blood genomic DNA and use of kit |
| CN103966205A (en) * | 2014-05-28 | 2014-08-06 | 谭晓刚 | Method for extracting ribonucleic acid from blood |
| CN106434894A (en) * | 2016-08-30 | 2017-02-22 | 江苏省弗泰生物科技有限公司 | Detection method for post-transplantation chimera |
| CN107245486A (en) * | 2017-06-21 | 2017-10-13 | 长沙金域医学检验所有限公司 | Poba gene group DNA extraction method |
| CN107988210A (en) * | 2018-01-09 | 2018-05-04 | 厦门基源医疗科技有限公司 | A kind of method of rapid extraction blood and buccal swab genomic DNA |
| CN110747255B (en) * | 2019-11-27 | 2023-08-22 | 郑州安图生物工程股份有限公司 | Pretreatment reagent and method for whole blood sample for nucleic acid detection |
| CN112725334B (en) * | 2021-02-23 | 2023-03-17 | 山东思科捷生物技术有限公司 | Cell RNA rapid extraction kit and RNA extraction method |
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| US20050142565A1 (en) * | 2003-12-30 | 2005-06-30 | Agency For Science, Technology And Research | Nucleic acid purification chip |
| WO2006067572A2 (en) * | 2004-12-14 | 2006-06-29 | University Of The Witwatersrand, Johannesburg | A method for diagnosing and monitoring cellular reservoirs of disease |
| CN101058595A (en) * | 2007-06-01 | 2007-10-24 | 吉林敖东药业集团延吉股份有限公司 | Ibonucleic acid, and preparation method and application thereof |
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| US20050142565A1 (en) * | 2003-12-30 | 2005-06-30 | Agency For Science, Technology And Research | Nucleic acid purification chip |
| WO2006067572A2 (en) * | 2004-12-14 | 2006-06-29 | University Of The Witwatersrand, Johannesburg | A method for diagnosing and monitoring cellular reservoirs of disease |
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