ⅠNacide ucleicintroduction
Acide nucleique igabanyijemo aside deoxyribonucleic (ADN) na aside ribonucleic (RNA), muri yo RNA igabanyijemo ribosomal RNA (rRNA), intumwa RNA (mRNA) no kohereza RNA (tRNA) ukurikije imirimo yayo itandukanye.ADN yibanda cyane muri nucleus, mitochondria na chloroform, naho RNA ikwirakwizwa cyane muri cytoplazme.Nka shingiro ryibintu byerekana imiterere ya gene, gukuramo aside nucleic bigira uruhare runini mubushakashatsi bwibinyabuzima bya molekuline no gusuzuma indwara ya molekile.Kwibanda no kwera kwa acide nucleic bizagira ingaruka kuri PCR ikurikiraho, ikurikiranye, kubaka vector, igogorwa rya enzyme nubundi bushakashatsi.
Ⅱ Gukuramo aside nucleic nuburyo bwo kweza
En Uburyo bwo gukuramo fenol / chloroform
Gukuramo Fenol / chloroform nuburyo bwa kera bwo gukuramo ADN, bukoresha cyane cyane ibishishwa bibiri bitandukanye bivura ingero, bigashonga aside nucleique ishingiye kuri ADN mugice cyamazi, lipide mugice kama, na proteyine hagati yibyiciro byombi.Ubu buryo bufite ibyiza byigiciro gito, ubuziranenge bwinshi ningaruka nziza.Ingaruka ni imikorere igoye kandi igihe kirekire.
Method Uburyo bwa Trizol
Uburyo bwa Trizol nuburyo bwa kera bwo gukuramo RNA.Uburyo bwa Trizol bugabanijwemo icyiciro cyamazi nicyiciro kama nyuma ya centrifugation hamwe na chloroform, aho RNA ishonga mugice cyamazi, icyiciro cyamazi cyimurirwa mumiyoboro mishya ya EP, imvura iboneka nyuma yo kongeramo isopropanol, hanyuma isukura Ethanol.Ubu buryo bukwiranye no gukuramo RNA mu nyamaswa, selile na bagiteri.
Method Uburyo bwo kweza inkingi
Uburyo bwo kweza inkingi ya Centrifuge burashobora kwamamaza ADN binyuze mubikoresho byihariye bya silicon matrix adsorption, mugihe RNA na proteyine bishobora kugenda neza, hanyuma ugakoresha umunyu mwinshi PH kugirango uhuze aside nucleic, umunyu muke wa PH ukuraho agaciro kugirango utandukanye kandi usukure aside nucleic.Ibyiza nibyiza byo kwezwa cyane, gutuza cyane, ntagikenewe kumashanyarazi, nigiciro gito.Ingaruka ni uko igomba gushyirwaho intambwe ku yindi, ibikorwa byinshi.
Method Uburyo bwo gukoresha amashanyarazi
Uburyo bw'amasaro ya magnetique ni ugucamo ibice by'utugingo ngengabuzima tunyuze muri lysate, kurekura aside nucleic muri sample, hanyuma molekile ya acide nucleique ikamenyeshwa cyane hejuru yisaro rya magneti, mugihe hasigaye umwanda nka proteyine nisukari. amazi.Binyuze mu ntambwe zo gutandukanya ingirabuzimafatizo, isaro ya magnetique ihuza aside nucleique, gukaraba aside nucleic, gukuramo aside nucleique, nibindi, aside nucleic irangije kuboneka.Ibyiza nibikorwa byoroshye nigihe gito cyo gukoresha, udakeneye intambwe ya centrifugation.Ifite tekinike nkeya kandi irashobora kumenya gukora byikora kandi rusange.Ihuriro ryihariye ryamasaro ya magnetique na acide nucleic ituma acide nucleic yakuwe hamwe nibitekerezo byinshi kandi byera.Ikibi nuko igiciro cyisoko kiriho ubu gihenze.
⑤ Ubundi buryo
Usibye uburyo bune buvuzwe haruguru, hariho guteka gutetse, uburyo bwumunyu mwinshi, uburyo bwa anionic detergent, uburyo bwa ultrasonic nuburyo bwa enzymatique, nibindi.
Ⅲ Ubwoko bwo gukuramo aside nucleic
Foregene ifite urubuga rwa mbere rwambere rwa PCR rwibanze, inkingi ebyiri RNA Isolation (ADN yonyine + RNA gusa).Ibicuruzwa byingenzi birimo ibikoresho byo kwigunga bya ADN / RNA, PCR na Direct PCR reagents ya molekile ya laboratoire.
Gukuramo RNA yose
Ingero zose zo gukuramo RNA zirimo amaraso, selile, ingirangingo zinyamanswa, ibimera, virusi, nibindi. clone ya molekulari, Akadomo hamwe nubundi bushakashatsi.
Foregene ifitanye isanoIbikoresho bya RNA
Amatungo Yuzuye ya RNA Yigunga--Byihuse kandi neza gukuramo RNA-yuzuye kandi yuzuye-yuzuye muri RNA inyama zinyamaswa zitandukanye.
Akagari Igiteranyo cya RNA Igikoresho cyo kwigunga--RNA isukuye cyane kandi yujuje ubuziranenge RNA irashobora kuboneka mumasoko atandukanye yimico muminota 11.
Tera Igiteranyo Cyuzuye cya RNA--Kuramo vuba RNA yuzuye yuzuye murwego rwibihingwa bifite polysaccharide nkeya hamwe na polifenol.
Virusi RNA Igikoresho cyo kwigunga--Gutandukanya kandi usukure virusi ya RNA mubitegererezo nka plasma, serumu, amazi yumubiri utagira selile hamwe numuco ndengakamere.
Gukuramo ADN
Ingero zo gukuramo ADN zirimo ubutaka, umwanda, amaraso, selile, inyama zinyamanswa, ibimera, virusi, nibindi. Gukuramo ADN ya genomic birashobora gukoreshwa mugusya enzyme, kubaka isomero rya ADN, PCR, gutegura antibody, isesengura rya blot yo mu burengerazuba, isesengura rya gene, hejuru -ibisubizo bikurikirana hamwe nubundi bushakashatsi.
Foregene ifitanye isanoIbikoresho bya ADN byo kwigunga
Inyamanswa yinyamanswa ADN yigunga- Gukuramo vuba no kweza ADN genomic biva ahantu henshi, nk'inyama z'inyamaswa, selile, nibindi.
Amaraso ADN Midi Kit (1-5ml)--Byiza vuba ADN ya genomic yo mu rwego rwo hejuru mumaraso ya anticagulée (1-5ml).
Buccal Swab / Ikarita ya FTA Ikarita yo kwigunga--Byihuse usukure ADN yo mu rwego rwohejuru ya ADN ya buccal swab / Ikarita ya FTA.
Tera ibikoresho bya ADN byo kwigunga--Kora neza kandi ubone ADN ya genomic yo mu rwego rwo hejuru uhereye ku bimera (harimo polysaccharide na polifenol)
Gukuramo plasmid
Plasmid ni ubwoko bwa molekile ntoya izenguruka ADN mu ngirabuzimafatizo, ikaba itwara abantu benshi mu kongera kwiyubaka kwa ADN.Uburyo bwo gukuramo plasmid ni ugukuraho RNA, gutandukanya plasmide na ADN ya bagiteri ya ADN, no kuvanaho poroteyine nindi myanda kugirango ubone plasmide isa neza.
Mini Mini Kit--Byihuse usukure ADN yo mu rwego rwohejuru ya plasmide ya bacteri zahinduwe kubushakashatsi bwibinyabuzima busanzwe bwa molekuline nko guhinduka no gusya kwa enzyme.
④ Ubundi bwoko bwo kuvoma, gukuramo miRNA, nibindi.
Inyamaswa miRNA Igikoresho cyo kwigunga--Byihuse kandi e ffi byoroshye gukuramo uduce duto twa RNA ya 20-200nt miRNA, siRNA, snRNA mubice bitandukanye byinyamanswa na selile.
Ⅳ Ibisabwa kugirango acide nucleic acide nibisubizo byo kwezwas
① Kugenzura ubusugire bwimiterere yibanze ya acide nucleic.
Kugabanya intungamubiri za poroteyine, isukari, lipide nizindi macromolecules
Should Ntihakagombye kubaho ibishishwa cyangwa ibinyabuzima byinshi bya ion bishobora kubuza enzyme mu byitegererezo bya aside nucleique.
④ RNA hamwe na aside nucleic yanduye igomba kuvaho mugihe ikuramo ADN, nibindi.
Igihe cyo kohereza: Ugushyingo-24-2022
