ⅠNʻakika ucleicihoʻolauna
Hoʻokaʻawale ʻia ka waikawa nucleic i ka waikawa deoxyribonucleic (DNA) a me ka waikawa ribonucleic (RNA), ma waena o ia RNA i māhele ʻia i ribosomal RNA (rRNA), messenger RNA (mRNA) a me ka hoʻololi ʻana i ka RNA (tRNA) e like me kāna mau hana like ʻole.Hoʻopili nui ʻia ka DNA i loko o ka nucleus, mitochondria a me chloroform, ʻoiai ʻo RNA ka mea i hoʻohele nui ʻia i ka cytoplasm.Ma ke ʻano kumu kumu o ka hōʻike ʻana, he hana koʻikoʻi ka ʻili nucleic acid i ka noiʻi biology molecular a me ka maʻi molecular clinical.E pili pono ana ka neʻe ʻana a me ka maʻemaʻe o ka ʻili o ka nucleic acid i ka PCR aʻe, ka hoʻonohonoho ʻana, ka hana vector, ka ʻeli ʻana o ka enzyme a me nā hoʻokolohua ʻē aʻe.
Ⅱ Nucleic acid extraction and purification method
① ʻO ke ʻano unuhi ʻana o Phenol/chloroform
ʻO ka unuhi ʻana o Phenol/chloroform he ʻano hana maʻamau no ka unuhi ʻana i ka DNA, ka mea nui e hoʻohana i ʻelua mau mea hoʻoheheʻe organik e mālama i nā laʻana, ka hoʻoheheʻe ʻana i ka waikawa nucleic i hoʻokumu ʻia i ka DNA i loko o ka pae wai, nā lipids i ka pae organik, a me nā protein ma waena o nā pae ʻelua.Loaʻa i kēia ʻano ka maikaʻi o ke kumukūʻai haʻahaʻa, ka maʻemaʻe kiʻekiʻe a me ka hopena maikaʻi.ʻO nā hemahema he hana paʻakikī a lōʻihi ka manawa.
② Keʻano Trizol
ʻO ke ʻano Trizol kahi ʻano maʻamau no ka unuhi ʻana o RNA.Hoʻokaʻawale ʻia ke ʻano Trizol i ka aqueous phase a me ka organic phase ma hope o ka centrifugation me ka chloroform, kahi i hoʻoheheʻe ʻia ai ka RNA i ka pae wai, ua hoʻoneʻe ʻia ka pae wai i kahi paipu EP hou, loaʻa ka ua ma hope o ka hoʻohui ʻana i ka isopropanol, a laila hoʻomaʻemaʻe ethanol.He kūpono kēia ʻano no ka unuhi ʻana i ka RNA mai nā ʻiʻo holoholona, nā cell a me ka bacteria.
③ ʻano hoʻomaʻemaʻe kolamu Centrifugal
Centrifuge kolamu hoʻomaʻemaʻe ala hiki adsorb DNA kiko'ī ma kūikawā silicon matrix adsorption mea, oiai RNA a me ka protein hiki ke hele smoothly, a laila, hoʻohana kiʻekiʻe paakai haʻahaʻa PH e hui nucleicʻakika, haʻahaʻa paʻakai kiʻekiʻe PH waiwai elution e hookaawale a hoomaemae nucleic waikawa.ʻO nā pōmaikaʻi he kiʻekiʻe ka hoʻomaʻemaʻe hoʻomaʻemaʻe, kūpaʻa kiʻekiʻe, ʻaʻohe pono no ka solvent organik, a me ke kumu kūʻai haʻahaʻa.ʻO ka hemahema ʻo ia ka mea pono e centrifuged i kēlā me kēia pae, ʻoi aku ka nui o ka hana.
④ ʻO ke ʻano o nā lei makeke
ʻO ke ʻano o ka peʻa magnetic ka hoʻokaʻawale ʻana i ka laʻana o ka cell tissue ma o ka lysate, hoʻokuʻu i ka nucleic acid i loko o ka hāpana, a laila hoʻopili pono ʻia nā molekala nucleic acid ma ka ʻili o ka peʻa magnetic, aʻo nā haumia e like me nā protein a me nā sugars e waiho ʻia i loko. ka wai.Ma o nā ʻanuʻu o ka hoʻokaʻawale ʻana o ka ʻiʻo cell, hoʻopaʻa ʻia ka bead magnetic me ka nucleic acid, ka holoi ʻana i ka wai nucleic, ka wai nucleic elution, a me nā mea ʻē aʻe, loaʻa ka waika nucleic maʻemaʻe.ʻO nā pōmaikaʻi he hana maʻalahi a me ka hoʻohana manawa pōkole, me ka ʻole o ka centrifugation step.Loaʻa iā ia nā koi ʻenehana haʻahaʻa a hiki ke hoʻomaopopo i ka hana aunoa a me ka nui.ʻO ka hui pū ʻana o ka magnetic bead a me ka nucleic acid e hana i ka ʻakika nucleic i unuhi ʻia me ke kiʻekiʻe a me ka maʻemaʻe.ʻO ka hemahema ʻo ia ke kumukūʻai o ka mākeke o kēia manawa.
⑤ Nā hana ʻē aʻe
Ma waho aʻe o nā ʻano ʻehā i luna aʻe, aia ka paila paila, ke ʻano paʻakai paʻakai, ke ʻano detergent anionic, ke ʻano ultrasonic a me ke ʻano enzymatic, etc.
Ⅲ Ke ʻano o ka ʻili o ka waika nucleic
Loaʻa iā Foregene ka papa kuhikuhi PCR alakaʻi honua, ʻelua kolamu RNA Isolation platform (DNA-wale + RNA wale nō).Aia nā huahana nui i nā pahu hoʻokaʻawale DNA/RNA, PCR a me Direct PCR reagents molecular lab reagents series.
① Huina RNA unuhi
ʻO ka nui o nā hōʻailona hōʻiliʻili RNA e loaʻa i ke koko, nā pūnaewele, nā ʻiʻo holoholona, nā mea kanu, nā virus, a me nā mea ʻē aʻe. molecular cloning, Dot Blot a me nā hoʻokolohua ʻē aʻe.
Foregene piliRNA Isolation Kit
Huina Hoʻokaʻawale RNA Holoholona--Hoʻopau wikiwiki a maikaʻi hoʻi i ka RNA holoʻokoʻa kiʻekiʻe-maʻemaʻe a kiʻekiʻe mai nā ʻiʻo holoholona like ʻole.
Kelepona Huina RNA Isolation Kit--Hiki ke loaʻa ka RNA piha hoʻomaʻemaʻe kiʻekiʻe a kiʻekiʻe mai nā ʻano moʻomeheu like ʻole i 11 mau minuke.
Mea Kanu Huina RNA Isolation Kit--Wehe koke i ka RNA holoʻokoʻa kiʻekiʻe mai nā mea kanu me ka polysaccharide haʻahaʻa a me ka polyphenol content.
Kiko Hoʻokaʻawale RNA Viral--E hoʻokaʻawale wikiwiki a hoʻomaʻemaʻe i ka RNA viral mai nā laʻana e like me ka plasma, serum, nā wai kino ʻole o ke kino a me nā supernatants moʻomeheu cell.
② Ka unuhi ʻana o Genomic DNA
ʻO nā hōʻailona hōʻiliʻili DNA Genomic e pili ana i ka lepo, feces, koko, cell, nā ʻiʻo holoholona, nā mea kanu, nā maʻi, a me nā mea ʻē aʻe. -ke kaʻina hana throughput a me nā hoʻokolohua ʻē aʻe.
Foregene piliNā pahu hoʻokaʻawale DNA
Kiko Hoʻokaʻawale DNA Tissue holoholona--Ka unuhi wikiwiki a me ka hoʻomaʻemaʻe ʻana i ka DNA genomic mai nā kumu he nui, e like me nā ʻiʻo holoholona, nā cell, etc.
Koko DNA Midi Kit (1-5ml)--Hoʻomaʻemaʻe wikiwiki i ka DNA genomic kiʻekiʻe mai ke koko anticoagulated (1-5ml).
Buccal Swab/FTA Kāleka DNA Isolation Kit--E hoʻomaʻemaʻe wikiwiki i ka DNA genomic kiʻekiʻe mai nā laʻana buccal swab/FTA Card.
Mea hoʻokaʻawale DNA mea kanu--E hoʻomaʻemaʻe wikiwiki a loaʻa ka DNA genomic kiʻekiʻe mai nā laʻana mea kanu (me nā polysaccharides a me nā laʻana mea kanu polyphenol)
③ Ka unuhi ʻana i ka plasmid
ʻO Plasmid kahi ʻano DNA mole liʻiliʻi liʻiliʻi i loko o nā pūnaewele, kahi mea lawe maʻamau no ka hui hou ʻana o DNA.ʻO ke ʻano o ka unuhi ʻana i ka plasmid ka wehe ʻana i ka RNA, hoʻokaʻawale i ka plasmid mai ka DNA genomic bacteria, a wehe i ka protein a me nā mea haumia ʻē aʻe e loaʻa ai ka plasmid maʻemaʻe.
Puke Mini Plasmid--Hoʻomaʻemaʻe wikiwiki i ka DNA plasmid kiʻekiʻe mai nā hua bacteria i hoʻololi ʻia no nā hoʻokolohua molecular biology maʻamau e like me ka hoʻololi ʻana a me ka digestion enzyme.
④ ʻO nā ʻano unuhi ʻē aʻe, miRNA extraction, etc.
Kiko hoʻokaʻawale miRNA holoholona--E wehe koke i nā ʻāpana RNA liʻiliʻi o 20-200nt miRNA, siRNA, snRNA mai nā ʻiʻo holoholona a me nā cell.
Ⅳ Nā koi no ka unuhi ʻana o ka waika nucleic a me ka hopena hoʻomaʻemaʻes
① No ka hōʻoia ʻana i ka pono o ka hana mua o ka waikawa nucleic.
② E hōʻemi i ka hoʻopili ʻana o nā protein, sugars, lipids a me nā macromolecules ʻē aʻe
③ ʻAʻohe mea hoʻoheheʻe organik a i ʻole ka nui o nā ion metala e hiki ke kāohi i ka enzyme i loko o nā ʻāpana nucleic acid.
④ Pono e hoʻopau ʻia ka RNA a me nā mea haumia nucleic acid i ka wā e unuhi ai i ka DNA, a me ka hope.
Ka manawa hoʻouna: Nov-24-2022
