I-Nucleic acid yahlulwe ibe yi-deoxyribonucleic acid (DNA) kunye ne-ribonucleic acid (RNA), phakathi kwayo i-RNA inokwahlulwa ibe yi-ribosomal RNA (rRNA), isithunywa RNA (mRNA) kwaye idlulisele i-RNA (tRNA) ngokwemisebenzi eyahlukeneyo.

I-DNA igxile kakhulu kwi-nucleus, mitochondria kunye ne-chloroplasts, ngelixa i-RNA isasazwa kakhulu kwi-cytoplasm.

Ngenxa yokuba iziseko ze-purine kunye neziseko ze-pyrimidine ziye zadibanisa iibhondi eziphindwe kabini kwi-nucleic acids, i-nucleic acids ineempawu zokufunxa i-ultraviolet. Ukufunxwa kwe-ultraviolet ye-DNA yetyuwa yesodium ijikeleze i-260nm, kwaye ukufunxa kwayo kubonakaliswa njenge-A260, kwaye ikwindawo yokufunxa kwi-230nm, ngoko ke i-ultraviolet spectroscopy ingasetyenziswa. I-Nucleic acids inqunywe ngokobungakanani kunye nekhwalithi yi-luminometer.

I-Nucleic acids zi-ampholytes, ezilingana ne-polyacids. I-Nucleic acids inokuhlukaniswa kwi-anion ngokusebenzisa i-neutral okanye i-alkaline buffers, kwaye ifakwe kwintsimi yombane ukuya kwi-anode. Lo ngumgaqo we-electrophoresis.

I-Nucleic acid extraction kunye nemigaqo yokucoca kunye neemfuno

1. Qinisekisa ukunyaniseka kwesakhiwo sokuqala se-nucleic acid

2. Ukuphelisa ukungcoliseka kwezinye iimolekyuli (ezifana nokungabandakanyi ukuphazamiseka kwe-RNA xa kutsalwa iDNA)

3. Akufuneki kubekho i-solvents eziphilayo kunye nokugxilwa okuphezulu kwee-ion zetsimbi ezivimbela i-enzymes kwiisampuli ze-nucleic acid.

4. Ukunciphisa izinto ze-macromolecular ezifana neeprotheni, iipolysaccharides kunye neelipids kangangoko kunokwenzeka.

I-Nucleic acid extraction kunye nendlela yokucoca

1. Phenol/chloroform extraction method

Yaqanjwa ngo-1956. Emva kokunyanga iseli ezaphukileyo zolwelo okanye izicubu homogenate ngephenol/chloroform, amacandelo nucleic acid, ikakhulu DNA, inyibilika kwisigaba aqueous, lipids ikakhulu kwisigaba eziphilayo, kwaye iiproteni zibekwe phakathi ezimbini. izigaba.

2. Ukuna kotywala

I-Ethanol inokuphelisa umaleko we-hydration we-nucleic acid kwaye iveze iqela le-phosphate ehlawuliswa kakubi, kwaye ii-ion ezichajiwe kakuhle ezifana ne-NA⋯⋯ zinokudityaniswa neqela le-phosphate ukwenza imvula.

3. Indlela yekholamu yeChromatographic

Ngokusebenzisa izixhobo ezikhethekileyo ze-silica-based adsorption, i-DNA inokubhengezwa ngokuthe ngqo, ngelixa i-RNA kunye neprotheni inokudlula ngokutyibilikayo, kwaye emva koko isebenzise ityuwa ephezulu kunye ne-pH ephantsi ukubopha i-nucleic acid, kunye ne-elute ngetyuwa ephantsi kunye ne-pH ephezulu yokwahlula kunye nokucoca i-nucleic. iasidi.

4. Indlela yealkali yokuqhekeka kweThermal

Ukutsalwa kwealkali ikakhulu kusebenzisa umahluko we-topological phakathi kwe-plasmids esetyhula evaliweyo kunye ne-chromatin emgqeni ukuze izahlule. Ngaphantsi kweemeko ze-alkaline, iiprotheyini ze-denatured zinyibilika.

5. Indlela yokubilisa ipyrolysis

Isisombululo se-DNA siphathwa ngobushushu ukuze sithathe inzuzo kwiipropathi ze-DNA ze-linear molecules ukwahlula amaqhekeza e-DNA kwi-precipitate eyenziwe ngama-proteins e-denatured kunye ne-debris yeselula nge-centrifugation.

6. Indlela yamaso eNanomagnetic

Ukusebenzisa i-nanotechnology ukuphucula kunye nokuguqula umphezulu we-superparamagnetic nanoparticles, i-superparamagnetic silicon oxide nano-magnetic beads ilungiselelwe. Amaso kazibuthe anokubona ngokuthe ngqo kwaye abophe ngokufanelekileyo kwiimolekyuli ze-nucleic acid kujongano oluncinci. Ukusebenzisa iipropathi ze-superparamagnetic ze-silica nanospheres, phantsi kwesenzo se-Chaotropic salts (i-guanidine hydrochloride, i-guanidine isothiocyanate, njl.) kunye nentsimi yangaphandle yamagnetic, i-DNA kunye ne-RNA yahlukaniswa negazi, izicubu zezilwanyana, ukutya, i-pathogenic microorganisms kunye nezinye iisampuli.