Nzira dzezvemakemikari yekutsvakurudza uye kushandisa kwavo

Nzira dzekutsvakurudza dzezvemakemikari dzinoita basa rinokosha mukurapa kwemazuva ano, mhirizhonga uye biology. Vongai nekufambira mberi mumunda weDNA neRNA zvidzidzo, munhu anokwanisa kudzidza ma genome ezvisikwa, ona kuti causative agent wechirwere ichi, anoziva yaidikanwa nucleic acid muzanganiswa yeacid, nezvimwewo.

Zvinyorwa-maitiro ezvokuongorora. Chii ichocho?

Kudzoka mumakore makumi manomwe nemakumi mapfumbamwe, masayendisiti akakwanisa kugadzirisa gome revanhu kwekutanga . Ichi chiitiko chakakonzera kukurudzira kwekugadzirwa kwema genetic engineering uye molecule biology. Kuongorora kwezvinhu zveDNA neRNA kwakaita kuti ikozvino zvigone kushandiswa idzi nucleic acids kuti uongorore chirwere, kudzidza majini.

Kugadzirira kweDNA neRNA

Nzira dzezvemakemikari yekuongororwa dzinoda kuvepo kwezvinhu zvakatangira: kazhinji ivo ndeve nucleic acids. Kune nzira dzakawanda dzekuparadzanisa zvinhu izvi kubva kumasero ezvisikwa zvipenyu. Chimwe nechimwe chazvo chine zvigaro zvayo uye zvisingabatsiri, uye izvi zvinofanirwa kuiswa mukutarisa pakusarudza nzira yekuparadzanisa nucleic acids muchimiro chakanaka.

1. Kugadzirira kweDNA maererano naMarmur. Iyo nzira inosanganisira mukurapa musanganiswa wezvinhu nemishonga inodhakwa, semugumisiro wei DNA yakachena inogadzirisa. Kuvhiringidza kweyi nzira ndiyo kushandiswa kwezvinhu zvinotyisa: phenol uye chloroform.

2. DNA kusanganiswa maererano neBoom. Chinhu chinoshandiswa pano chinonzi guanidine thiocyanate (GuSCN). Inokurudzira kunyura kwe deoxyribonucleic acid pane michina yakasiyana, iyo inogona kuendeswa mberi nayo nechigadzirwa chakakosha. Zvisinei, GuSCN inhibitor yePTC, uye kunyange chikamu chiduku chayo, chakabatanidzwa muDNA yakanyanyisa, inogona kukanganisa nzira ye polymerase chain reaction, iyo inoita basa rinokosha mubasa ne nucleic acids.

3. Kuparara kwezvisina kunaka. Iyo nzira inopesana neiyo yapfuura mune kuti mamolekemu e deoxyribonucleic acid ivo inogadziriswa, asi kusvibiswa. Kuti uite izvi, shandisa ion exchangers. Izvo zvinotadza ndezvokuti haisi zvinhu zvose zvinogona kugadzirisa.

4. Kuongorora maziso. Iyi nzira inoshandiswa mune izvo zviitiko apo ruzivo rwakananga pamusoro pekuumbwa kwema molecule eDNA hazvidiwi, asi zvakakosha kuwana imwe tsananguro yedata. Izvi zvinotsanangurwa nenyaya yokuti chimiro che nucleic acid chinogona kukuvadzwa nekurapa ne detergents, kunyanya, nealkalis.

Urongwa hwekutsvakurudza nzira

Maitiro ose emakemikari ekutsvakurudza akaparadzaniswa kuva mapoka makuru matatu:

1. Amplification (kushandiswa kwemhando dzakasiyana-siyana dze enzymes). Izvi zvinosanganisira PCR-polymerase chain reaction, iyo inoita basa guru mumaitiro akawanda ekuongororwa.

2. Asina-appellation. Iri boka re nzira rinowirirana zvakananga nebasa rekuvhenganiswa kwe nucleic acids. Mienzaniso mitatu yemhando yekudzima, in situ hybridization, nezvimwewo.

3. Nzira dzakabva pakuzivikanwa kwechiratidzo kubva kune probe molecule inosunga kune imwe DNA kana RNA probe. Muenzaniso urongwa hwehuwandu hweHybride Capture System (hc2).

Inymei inogona kushandiswa mumakemikari ezvinyorwa zvesayenzi

Nzira dzakawanda dzekuongororwa kwemakemikari dzinosanganisira kushandiswa kwemhando dzakawanda dze enzymes. Pasi apa ndiyo inowanzoshandiswa:

1. Restrictase - "kucheka" damolekemu reDNA muzvikamu zvinodiwa.

2. DNA polymerase - inogadzira kaviri-yakanyunguduka mamolekemu ye deoxyribonucleic acid.

3. Reverse transcriptase (revertase) - inoshandiswa pakugadzira DNA pamutambo weRNA.

4. DNA ligase - inotungamirira pakuumbwa kwephosphodiester zvisungo pakati pe nucleotides.

5. Exonuclease - inobvisa nucleotides kubva kumagumo ekupedzisira e molecule ye deoxyribonucleic acid.

PCR ndiyo nzira huru yekuwedzera DNA

Polymerase chain reaction (PCR) inoshandiswa zvakashandiswa mumazuva ano masayendisiti ezvinyorwa. Iyi ndiyo nzira iyo inhamba yakawanda yemakopi inogona kuwanikwa kubva kune imwe DNA molecule (kukurisa mamolekemu).

Zvinonyanya basa zvePCR ndezvi:

- Kuziva zvirwere;

Kugadzirisa DNA, majini.

Iyo polymerase chain reaction inoda zvinotevera: dNA yakatanga DNA molecules, DNA polymerase inotonhora (Taq kana Pfu), deoxyribonucleotide phosphates (inotora nitrogen mabhesi), inotanga (2 primers pamusana weDNA molecule) uye iyo inonzi buffer system iyo, iyo inogona kuita Pamusoro pezvinoitika zvose.

PCR inosanganisira matanho matatu: kuchenesa, kushandiswa kwezvipfeko uye kuwedzera.

1. Denaturation. Pakupisa kwe 94-95 degrees Celsius, kuputsa kwehutano hwehydrogen pakati pemadziro maviri eDNA inoenderera mberi, uye somugumisiro tinowana ma-molecules maviri.

2. Annealing of the primers. Pachando choupfu cha 50-60 degrees Celsius, zvinyorwa zvinonamatirwa pamagumo e-single-stranded nucleic acid molecules maererano nerudzi rwekubatanidzwa.

3. Kusarudzwa. Pachishambiso che 72 degrees, kufanana kwekunda kwemaviri emakemikoroni e deoxyribonucleic acid inowanikwa.

DNA Sequencing

Nzira dzezvemakemikari dzinowanzodikanwa kuziva ruzivo rwekugadzirisa kweiucleotides mune imwe molecule ye deoxyribonucleic acid. Kuchengetedza kunoshandiswa kutsanangura genetic code . Zviratidzo zvemasero zvezveramangwana zvichabva pane ruzivo rwakagadziriswa mukugadzirisa kuenzanirana kwemunhu.

Mhando dzekutevera dzinotevera dzinozivikanwa:

• Kutarisa naMaamam-Gilbert;
• Kuenzanisa naSanger;
• Pyrosequencing;
• Nanoporous sequencing.