Hoʻokaʻawale ʻia ka waikawa nucleic i ka waikawa deoxyribonucleic (DNA) a me ka waikawa ribonucleic (RNA), ma waena o ia mau mea hiki ke hoʻokaʻawale ʻia ʻo RNA i ribosomal RNA (rRNA), messenger RNA (mRNA) a me ka RNA transfer (tRNA) e like me nā hana like ʻole.

Hoʻopili nui ʻia ʻo DNA i loko o ka nucleus, mitochondria a me nā chloroplasts, ʻoiai ʻo RNA ka mea i māhele nui ʻia i ka cytoplasm.

No ka mea ua hoʻohui nā kumu purine a me nā kumu pyrimidine i nā paʻa pālua i nā waikawa nucleic, loaʻa i nā ʻakika nucleic nā ʻano o ka absorption ultraviolet. ʻO ka ultraviolet absorption o nā paʻakai sodium DNA ma kahi o 260nm, a ua hōʻike ʻia kona absorbance e like me A260, a aia ma ka absorption trough ma 230nm, no laila hiki ke hoʻohana ʻia ka spectroscopy ultraviolet. Hoʻoholo ʻia nā ʻakika nucleic i ka nui a me ka qualitatively e ka luminometer.

ʻO nā ʻakika nucleic he ampholytes, ua like ia me nā polyacids. Hiki ke hoʻokaʻawale ʻia nā ʻakika nucleic i nā anion me ka hoʻohana ʻana i nā pale kūʻokoʻa a i ʻole ka alkaline, a waiho ʻia i loko o ke kahua uila e neʻe ai i ka anode. ʻO kēia ke kumu o ka electrophoresis.

Nucleic acid extraction a me ka hoʻomaʻemaʻe kumu a me nā koi

1. E hōʻoia i ka paʻa o ka ʻakika nucleic kumu mua

2. Hoʻopau i ka haumia o nā molekole ʻē aʻe (e like me ka haʻalele ʻana i ka RNA interference i ka wā e unuhi ai i ka DNA)

3. ʻAʻole pono e loaʻa nā mea hoʻoheheʻe organik a me nā kiʻekiʻe kiʻekiʻe o nā iona metala e kāohi ai i nā enzymes i loko o nā ʻāpana nucleic acid.

4. E ho'ēmi i nā mea macromolecular e like me nā proteins, polysaccharides a me nā lipid e like me ka hiki.

Nucleic acid extraction and purification method

1. Phenol/chloroform kaʻina hana

Ua hana ʻia i ka makahiki 1956. Ma hope o ka mālama ʻana i ka wai i haki ʻia a i ʻole ka homogenate kiko me ka phenol/chloroform, ua hoʻoheheʻe ʻia nā ʻāpana nucleic acid, ka nui o ka DNA, i loko o ka aqueous phase, lipids ka hapa nui i ka organic phase, a aia nā protein i waena o nā mea ʻelua. nā māhele.

2. Ka ua waiʻona

Hiki i ka Ethanol ke hoʻopau i ka papa hydration o ka nucleic acid a hōʻike i ka pūʻulu phosphate i hoʻopiʻi ʻole ʻia, a hiki i nā ion i hoʻopaʻa maikaʻi ʻia e like me NA﹢ hiki ke hui pū me ka pūʻulu phosphate e hana i kahi eha.

3. Ke ʻano kolamu Chromatographic

Ma o ka silica-based adsorption material, hiki ke hoʻopili pono ʻia ka DNA, ʻoiai hiki i ka RNA a me ka protein ke hele mālie, a laila hoʻohana i ka paʻakai kiʻekiʻe a me ka pH haʻahaʻa e hoʻopaʻa ai i ka waikawa nucleic, a elute me ka paʻakai haʻahaʻa a me ka pH kiʻekiʻe e hoʻokaʻawale a hoʻomaʻemaʻe i ka nucleic. ʻakika.

4. Thermal māhā alkali ala

Hoʻohana maʻamau ka unuhi ʻana i ka alkaline i nā ʻokoʻa topological ma waena o nā plasmids pōʻai covalently i pani ʻia a me linear chromatin e hoʻokaʻawale iā lākou. Ma lalo o nā kūlana alkaline, hiki ke hoʻoheheʻe ʻia nā protein denatured.

5. Keʻano pyrolysis paila

Hoʻomaʻamaʻa ʻia ka hopena DNA e hoʻohana i nā waiwai o nā molekele DNA linear e hoʻokaʻawale i nā ʻāpana DNA mai ka precipitate i hana ʻia e nā protein denatured a me nā ʻōpala cellular e ka centrifugation.

6. ʻO ke ʻano o nā lei nanomagnetic

Ke hoʻohana nei i ka nanotechnology e hoʻomaikaʻi a hoʻololi i ka ʻili o nā nanoparticles superparamagnetic, ua hoʻomākaukau ʻia nā silicon oxide nano-magnetic beads superparamagnetic. Hiki ke hoʻomaopopo pono a hoʻopaʻa pono i nā molekala nucleic acid ma kahi interface microscopic. Ke hoʻohana nei i nā waiwai superparamagnetic o nā silica nanospheres, ma lalo o ka hana o nā paʻakai Chaotropic (guanidine hydrochloride, guanidine isothiocyanate, a me nā mea ʻē aʻe) a me kahi kahua magnetic waho, ua hoʻokaʻawale ʻia ʻo DNA a me RNA mai ke koko, nā holoholona holoholona, ​​​​meaʻai, nā microorganisms pathogenic a me nā mea hoʻohālike ʻē aʻe.