Nucleic acid extraction column method and principle

Nucleic acid ua vaevaeina i le deoxyribonucleic acid (DNA) ma le ribonucleic acid (RNA), lea e mafai ai ona vaevaeina RNA i le ribosomal RNA (rRNA), avefeau RNA (mRNA) ma le fesiitaiga RNA (tRNA) e tusa ai ma galuega eseese.

O le DNA o loʻo faʻapipiʻiina i totonu o le nucleus, mitochondria ma chloroplasts, aʻo RNA e masani ona tufatufaina i le cytoplasm.

Talu ai ona o faʻavae purine ma faʻavae pyrimidine ua faʻapipiʻi faʻalua faʻatasi i nucleic acids, o le nucleic acids o loʻo i ai uiga o le ultraviolet absorption.O le ultraviolet absorption o DNA sodium salts e tusa ma le 260nm, ma o lona absorbance o loʻo faʻaalia e pei o le A260, ma o loʻo i le absorption trough i le 230nm, o lea e mafai ai ona faʻaogaina le ultraviolet spectroscopy.Nucleic acids e fua fa'atatau ma fa'atatau e se luminometer.

Nucleic acids o ampholytes, e tutusa ma polyacids.Nucleic acids e mafai ona fa'amavae i anion e ala i le fa'aogaina o pa'u e le tutusa po'o le alkaline, ma tu'u i totonu o se eletise eletise e fa'asolo atu i le anode.O le mataupu faavae lea o le electrophoresis.

Nucleic acid extraction column method and principle

Nucleic acid extracting ma fa'amamāina ta'iala ma mana'oga

1. Faʻamautinoa le faʻamaoni o le faʻavae autu o le nucleic acid

2. Ave'ese le fa'aleagaina o isi molela'au (e pei o le le aofia ai o le RNA fa'alavelave pe a aveese le DNA)

3. E le tatau ona i ai ni solvents faʻaletino ma maualuga faʻatonuga o ion uʻamea e faʻalavelaveina enzymes i faʻataʻitaʻiga nucleic acid.

4. Faʻaitiitia mea faʻapitoa e pei o polotini, polysaccharides ma lipids i le tele e mafai ai.

Nucleic acid extracting and purification method

1. Fa'atosina ole phenol/chloroform

Na faia i le 1956. Ina ua maeʻa ona togafitia le sela gau vai poʻo le mea tutusa i le phenol / chloroform, o vaega o le nucleic acid, aemaise lava le DNA, e faʻamavaeina i le vaega vai, o le lipids e tele lava i le vaega o meaola, ma o loʻo i ai le polotini i le va o le lua. vaega.

2. Fa'asusu le ava malosi

E mafai e le ethanol ona fa'aumatia le vaega fa'asusu o le nucleic acid ma fa'aalia ai le vaega o le phosphate leaga, ma o ion e molia lelei e pei ole NA﹢ e mafai ona tu'u fa'atasi ma le vaega phosphate e fa'atupu ai se ma'a.

3. auala koluma Chromatographic

E ala i le mea faapitoa adsorption faavae silica, DNA e mafai ona faapitoa adsorbed, ao RNA ma polotini e mafai ona ui atu sologa lelei, ona faaaoga ai lea o le masima maualuga ma le pH maualalo e fusifusia ai le nucleic acid, ma elute i le masima maualalo ma le pH maualuga e vavae ese ai ma faamamaina nucleic. acid.

4. auala alkali ta'e vevela

Ole fa'aaogāina ole alkaline e fa'aaoga ai le eseesega i luga ole va ole plasmids lapotopoto fa'amuta ma le chromatin laina e vavae'ese ai.I lalo o tulaga alkaline, o polotini denatured e soluble.

5. Fa'apuna pyrolysis auala

Ole fofo ole DNA e fa'afefeteina e fa'aoga lelei ai meatotino ole laina ole DNA molelaula e tu'u'ese'ese ai vaega ole DNA mai le fa'afefete na faia e polotini denatured ma otaota feavea'i e ala ile centrifugation.

6. Nanomagnetic beads metotia

O le faʻaaogaina o le nanotechnology e faʻaleleia ma faʻaleleia ai le pito i luga ole superparamagnetic nanoparticles, superparamagnetic silicon oxide nano-magnetic beads ua saunia.O lopa mageta e mafai ona iloa ma'oti ma pipii lelei i molela'au nucleic acid i luga o se atina'e microscopic.O le faʻaaogaina o le superparamagnetic properties o silica nanospheres, i lalo o le gaioiga a Chaotropic salts (guanidine hydrochloride, guanidine isothiocyanate, ma isi) ma se fanua mageta i fafo, DNA ma RNA na vavae ese mai le toto, manu manu, meaʻai, microorganisms pathogenic ma isi faʻataʻitaʻiga .


Taimi meli: Mati-18-2022