| The nucleic acid is playing a very important role in the continuation ofthe living organisms and the process of evolution as the genetic informationcarriers of the living organisms. Now the study of nucleic acid includes thequalitative and quantitative research. In the quantitative study of mainapplication The technologies used in the the quantitative study of nucleic acidare the polymerase chain reaction (PCR) technology, high performance liquidchromatography (HPLC),the titration and digital PCR technology. Massspectrometry technology and infrared spectral analysis technology has beenapplied in the study of the nucleic acid qualitative. For the research of nucleicacid, this paper is as follows:1. The development of adenine and cytosine standard materialAdenine and cytosine of the bases of deoxyribonucleic acid (DNA) havesome weak alkali as a result of two kinds of bases contain amino groups.Thetwo bases were quantified by the titration in the paper. By the optimization oftitration agent type, concentration and the solvents type, the concentration of0.05mol/L high chlorine acid solution was selected as titrant and glacialacetic acid was selected as solvent in the research. Int he following study,using HPLC was used the research of the stability of two bases andMass spectrometry technology and infrared spectral analysis technology wereapplied in the study of the bases qualitative and also the ash content and thewater content of the sample were analysed. The results are shown in thefollowing after uncertainty analysi:the two standard material content ofadenine and cytosine were99.40±0.37%，99.50±1.07%.2. quantitative research of Lambda DNA by digital PCRDigital PCR technology is a new technology without the standard ofcalibration, which is used the research of nucleic acid because of theadvantages of low concentration, high accuracy. The experiment quantifiedlambda DNA samples by dPCR and evaluated the test uncertainty. Resultshows that lambda DNA was quantified as2.428μg with an uncertainty of0.1305(K=2).3. The analysis of SNP by MSAs a commonly analysis method,Mass spectrometry has been used in theresearch of single nucleotide polymorphisms (SNPS) with tshort time ofoperation, high throughput, high sensitivity. the single nucleotidepolymorphisms (SNPS) are analyzed by the MAILD-TOF in the research. Thehigh variable carrier of the mitochondrial DNA sequence HV2area wasstructured in the experiment and PCR amplification was carried out.In thefollowing research, single base extension reaction was expanded. Then thePCR products were purified by the purification methods of solid-phase extraction and magnetic beads method. The purification products wereanalysed by mass spectrometry.Through the comparison of the molecularweight difference of the extension product to get the base type. Three singlenucleotide polymorphisms (SNPS) were detected in the experiment and eachsites was detected by five times. Finally the experimental results are as thesame wite the theoretical results.4. the comparison of three purification methods in the research of sequencingThis study aimed to establish a kind of good purification method for PCRproducts in the subsequent sequencing reaction. short tandem repeat D2S441clips as the template, PCR amplification reactionw was implemented and thePCR products were analysed by Agilent2100Bioanalyzer.The purificationmethod for PCR product based on solid-phase extraction by96holes OasisHLB SPE plate and magnetic beads and purification kits were demonstrated.Final results show that the purification method of solid phase extraction isbetter than the purification kits method and magnetic beads method.5. Cooperation value and uncertainty analysis for transgenic RapeseedTOPAS19/2reference plasmid moleculeBy the feature of easy acquisition, low cost and short term, PlasmidReference Materials could be used for GMO quantifications. In thisexperiment, Real-time quantitative PCR was used to quantify GMO RapeseedTOPAS19/2collaboratively by7laboratories. The genome alternativeresearch, collaborative and fixed value research and the uncertainty research of transgenic rapeseed TOPAS19/2plasmid molecules were made by7collaborative laboratory. The collaborative laboratory results showed thatRapeseed TOPAS19/2was quantified as0.910with an uncertainty of0.013(K=2)。... |
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