ⅠNucleic acidifolasaga
Nucleic acid ua vaevaeina i le deoxyribonucleic acid (DNA) ma le ribonucleic acid (RNA), lea e vaevaeina ai le RNA i le ribosomal RNA(rRNA), avefeau RNA(mRNA) ma le fesiitaiga RNA(tRNA) e tusa ai ma ana galuega tauave eseese.O le DNA o loʻo faʻapipiʻiina i totonu o le nucleus, mitochondria ma chloroform, aʻo RNA e masani ona tufatufaina i le cytoplasm.I le avea ai ma faavae o le faʻamatalaga o kenera, o le faʻaaogaina o le nucleic acid e taua tele i suʻesuʻega o meaola mole mole ma suʻesuʻega mole mole.O le faʻatonuga ma le mama o le faʻaaogaina o le nucleic acid o le a aʻafia saʻo ai le PCR mulimuli ane, faʻasologa, fausiaina o vete, faʻamaʻi enzyme ma isi suʻega.
Ⅱ Nucleic acid extraction ma le fa'amamāina
① Fa'ata'ita'iga o le phenol/chloroform
Phenol/chloroform extraction ose auala masani mo DNA extraction, lea e masani ona faʻaogaina ni solvents eseese e lua e togafitia ai faʻataʻitaʻiga, faʻamavaeina le DNA faʻavae nucleic acid i le vai, lipids i le vaega faʻaola, ma polotini i le va o vaega e lua.O lenei metotia o loʻo i ai le lelei o le tau maualalo, maualuga le mama ma le lelei.O mea le lelei o le faʻaogaina faigata ma le umi o le taimi.
② Trizol auala
Trizol metotia o se auala masani mo le faʻaaogaina o le RNA.O le auala Trizol ua vaevaeina i le vaega vai ma le vaega faʻaletino pe a maeʻa le centrifugation ma le chloroform, lea e faʻamavaeina ai le RNA i le vaega aqueous, o le vaega aqueous e faʻafeiloaʻi i se EP fou tube, e maua le vaʻa pe a uma ona faʻaopopo le isopropanol, ona faʻamamaina ai lea o le ethanol.O lenei metotia e talafeagai mo le RNA fa'aoso mai aano o manu, sela ma siama.
③ auala fa'amamāina koluma centrifugal
Centrifuge koluma auala faamamaina e mafai ona adsorb DNA faapitoa e ala faapitoa silicon matrix adsorption mea, ao RNA ma polotini e mafai ona pasi sologa lelei, ona faaaoga ai lea o le masima maualuga maualalo PH e tuufaatasia ai le nucleic acid, masima maualalo maualuga PH elution e vavae ese ma faamamaina nucleic acid.O mea lelei e maualuga le faʻamamaina o le faʻamamaina, maualuga le mautu, leai se manaʻoga mo solvent organic, ma le tau maualalo.O le faʻaletonu e manaʻomia le centrifuged i lea laasaga ma lea laasaga, sili atu laasaga faʻagaioiga.
④ auala maneta lopa
O le auala o lopa mageta o le vaeluaina lea o le cell tissue sample e ala i le lysate, faʻamalolo le nucleic acid i totonu o le faʻataʻitaʻiga, ona faʻapipiʻi faapitoa ai lea o molelaʻau nucleic acid i luga o le pito i luga ole maneta, ae o mea leaga e pei o polotini ma suka o loʻo totoe i totonu. le vai.E ala i laʻasaga o le vaeluaga o le tino, maneta lopa fusifusia ma nucleic acid, nucleic acid fufulu, nucleic acid elution, ma isi, mama nucleic acid mulimuli ane maua.O mea lelei o le faʻaogaina faigofie ma le faʻaaogaina o le taimi puupuu, e aunoa ma le manaʻomia o le centrifugation laasaga.E maualalo manaoga faʻapitoa ma e mafai ona iloa otometi ma tele galuega.O le tu'ufa'atasiga fa'apitoa o le pulu mageta ma le nucleic acid e maua mai ai le nucleic acid ma le maualuga ma le mama.O le tulaga le lelei o le tau maketi o loʻo i ai nei e fai si taugata.
⑤ Isi metotia
I le faaopoopo atu i auala e fa o loʻo i luga, o loʻo i ai le faʻafefeteina o le faʻafefe, metotia masima faʻapitoa, metotia faʻamalo anionic, metotia ultrasonic ma le enzymatic metotia, ma isi.
Ⅲ Ituaiga o le su'ega nucleic acid
Foregene o loʻo i ai le taʻutaʻua taʻutaʻua o le PCR tuʻufaʻatasia i le lalolagi, faʻailoga lua RNA Isolation platform(DNA-naʻo + RNA naʻo ).O oloa autu e aofia ai pusa vavae DNA/RNA, PCR ma Direct PCR reagents molecular lab reagents series.
① Aofa'i RNA uma
Ole aofaʻi ole faʻataʻitaʻiga ole RNA e aofia ai le toto, sela, aano o manu, laʻau, viruses, ma isi. Ole mama maualuga ma le maualuga ole maualuga ole RNA e mafai ona maua e ala ile aofaʻi atoa ole RNA, lea e mafai ona faʻaaogaina ile RT-PCR, suʻesuʻega pu, faʻaliliuga in vitro, mole mole cloning, Dot Blot ma isi su'ega.
Foregene fesootaiRNA Isolation Kits
Manu Aofa'i RNA Isolation Kit--Fa'atopetope ma fa'alelei le fa'amama fa'amama ma le maualuga o le aofa'i atoa o le RNA mai aano o manu eseese.
Cell Total RNA Isolation Kit--E mafai ona maua le aofa'i atoa o le RNA fa'amama ma maualuga mai sela fa'aleaganu'u eseese ile 11 minute.
La'au Aofa'i RNA Isolation Kit--Fa'atopetope ona aveese le RNA atoa mai la'au fa'ata'ita'iga e maualalo le polysaccharide ma le polyphenol.
Viral RNA Isolation Kit--Fa'ate'a vave ma fa'amama RNA viral mai fa'ata'ita'iga e pei ole plasma, serum, su'a ole tino e leai se sela ma su'ega aganu'u.
② Su'ega DNA genomic
Genomic DNA faʻataʻitaʻiga faʻataʻitaʻiga e aofia ai le eleele, feces, toto, sela, aano o manu, laʻau, siama, ma isi. Genomic DNA extraction e mafai ona faʻaaogaina i le faʻaogaina o le enzyme, fausiaina o le faletusi DNA, PCR, sauniuniga antibody, Western blot hybridization analysis, gene chip, maualuga. - fa'asologa fa'asologa ma isi fa'ata'ita'iga.
Foregene fesootaiDNA Tu'uesea Pusa
Pusa Tu'uesea DNA Tissue Manu--Ave vave ma faʻamamāina le genomic DNA mai le tele o punaoa, e pei o aano o manu, sela, ma isi.
Toto DNA Midi Kit (1-5ml)- Fa'amama vave le DNA genomic sili ona lelei mai le toto fa'ato'a (1-5ml).
Buccal Swab/FTA Card DNA Isolation Kit--Fa'amama vave le DNA genomic sili ona lelei mai fa'ata'ita'iga o fa'ata'ita'iga o le swab/FTA Card.
La'au DNA Tu'uesea Pusa--Fa'amama vave ma maua le DNA genomic sili ona lelei mai la'au fa'ata'ita'iga (e aofia ai polysaccharides ma polyphenol la'au fa'ata'ita'iga)
③ Fa'apipi'i plasmid
O le Plasmid o se ituaiga o DNA mole laʻititi liʻo i totonu o sela, o se ave masani mo DNA recombination.O le auala o le plasmid extraction o le aveese lea o le RNA, vavae ese le plasmid mai le siama genomic DNA, ma aveese le polotini ma isi mea leaga e maua ai le plasmid mama.
General Plasmid Mini Kit- Fa'amama vave le DNA plasmid maualuga mai siama ua suia mo fa'ata'ita'iga masani o meaola mole mole e pei o le suiga ma le fa'aiina o le enzyme.
④ O isi ituaiga fa'atosina, miRNA extraction, etc.
Manu miRNA Tu'uesea Pusa--Vave vave ma lelei ona aveese fasi vaega RNA laiti o le 20-200nt miRNA, siRNA, snRNA mai aano o manu eseese ma sela.
Ⅳ Manaoga mo le su'eina o le nucleic acid ma le fa'amamāinas
① Faʻamautinoa le faʻamaoni o le fausaga muamua o le nucleic acid.
② Faʻaitiitia le faʻalavelaveina o polotini, suka, lipids ma isi macromolecules
③ E le tatau ona i ai se solvent organic poʻo le maualuga o le faʻaogaina o ion uʻamea e mafai ona faʻalavelaveina le enzyme i faʻataʻitaʻiga nucleic acid.
④ RNA ma isi faʻamaʻi nucleic acid e tatau ona faʻaumatia pe a aveese le DNA, ma le isi itu.
Taimi meli: Nov-24-2022
