ⅠNucleic acidigabatarwa
Nucleic acid ya kasu kashi deoxyribonucleic acid (DNA) da ribonucleic acid (RNA), daga cikinsu an raba RNA zuwa ribosomal RNA(rRNA), messenger RNA (mRNA) da kuma canja wurin RNA (tRNA) bisa ga ayyukansa daban-daban.DNA ya fi maida hankali ne a cikin tsakiya, mitochondria da chloroform, yayin da RNA ke rarrabawa a cikin cytoplasm.A matsayin ginshiƙi na bayanin kwayoyin halitta, cirewar acid nucleic yana taka muhimmiyar rawa a cikin binciken nazarin halittun kwayoyin halitta da ganewar ƙwayoyin ƙwayoyin cuta na asibiti.Mahimmanci da tsabtar hakar acid nucleic zai shafi kai tsaye PCR na gaba, jerin abubuwa, ginin vector, narkewar enzyme da sauran gwaje-gwaje.
Ⅱ Nucleic acid hakar da kuma hanyar tsarkakewa
① Hanyar cire phenol/chloroform
Phenol/chloroform hakar hanya ce ta gargajiya don hakar DNA, wanda galibi yana amfani da abubuwan kaushi daban-daban guda biyu don bi da samfuran, narkar da DNA tushen nucleic acid a cikin lokaci na ruwa, lipids a cikin tsarin kwayoyin halitta, da kuma sunadarai tsakanin matakan biyu.Wannan hanya tana da fa'idodi na ƙananan farashi, babban tsabta da tasiri mai kyau.Rashin lahani shine aiki mai rikitarwa da kuma dogon lokaci.
② Hanyar Trizol
Hanyar Trizol hanya ce ta gargajiya don hakar RNA.Hanyar Trizol ta kasu kashi kashi na ruwa da kuma kwayoyin halitta bayan centrifugation tare da chloroform, wanda aka narkar da RNA a cikin lokaci mai ruwa, an canza yanayin ruwa zuwa sabon bututun EP, ana samun hazo bayan ƙara isopropanol, sa'an nan kuma tsarkakewar ethanol.Wannan hanya ta dace da hakar RNA daga kyallen dabbobi, sel da ƙwayoyin cuta.
③ Hanyar tsarkakewa shafi na Centrifugal
Hanyar tsarkakewa shafi na Centrifuge na iya adsorb DNA ta musamman ta kayan tallan kayan siliki na musamman, yayin da RNA da furotin na iya wucewa lafiya, sannan amfani da babban gishiri low PH don haɗa acid nucleic, ƙarancin gishiri mai ƙimar darajar PH don ware da tsarkake acid nucleic.Abubuwan da ake amfani da su sune babban taro na tsarkakewa, babban kwanciyar hankali, babu buƙatar maganin kwayoyin halitta, da ƙananan farashi.Lalacewar shi ne cewa yana buƙatar a daidaita shi mataki-mataki, ƙarin matakan aiki.
④ Hanyar Magnetic beads
Hanyar Magnetic Beads ita ce raba samfurin tantanin halitta ta hanyar lysate, saki acid nucleic da ke cikin samfurin, sannan kuma kwayoyin nucleic acid suna daɗaɗa musamman a saman dutsen maganadisu, yayin da ƙazanta irin su proteins da sugars aka bar su a ciki. ruwa.Ta hanyar matakan rarrabuwar ƙwayoyin sel, igiyar maganadisu mai ɗaure tare da acid nucleic, wankin acid nucleic, elution nucleic acid, da sauransu, a ƙarshe an sami tsantsar nucleic acid.Abubuwan amfani sune aiki mai sauƙi da amfani da ɗan gajeren lokaci, ba tare da buƙatar matakin centrifugation ba.Yana da ƙananan buƙatun fasaha kuma yana iya gane aiki ta atomatik da taro.Haɗin ƙayyadaddun ƙayyadaddun ƙwayar maganadisu da acid nucleic yana sanya acid ɗin nucleic da aka fitar tare da babban taro da tsabta.Rashin hasara shine cewa farashin kasuwa na yanzu yana da tsada sosai.
⑤ Sauran hanyoyin
Baya ga hanyoyin huɗun da ke sama, akwai fashewar tafasa, hanyar gishiri mai mai da hankali, hanyar wankan anionic, hanyar ultrasonic da hanyar enzymatic, da sauransu.
Ⅲ Nau'in hakar acid nucleic
Foregene yana da babban dandamalin PCR na kai tsaye na duniya, dandamalin keɓewar RNA guda biyu (DNA-kawai + RNA kawai).Babban samfuran sun haɗa da kayan keɓewar DNA/RNA, PCR da Direct PCR reagents jerin reagents na kwayoyin halitta.
① Jimlar hakar RNA
Jimlar samfuran cirewar RNA sun haɗa da jini, sel, kyallen dabba, tsire-tsire, ƙwayoyin cuta, da dai sauransu. Ana iya samun tsafta mai girma da babban taro na jimlar RNA ta hanyar cirewar RNA gaba ɗaya, wanda za'a iya amfani dashi a cikin RT-PCR, binciken guntu, fassarar in vitro, cloning kwayoyin halitta, Dot Blot da sauran gwaje-gwaje.
Alamar farkoRNA Isolation Kits
Jimlar Dabbobin Keɓewar RNA--Cikin sauri da inganci cire tsafta mai inganci da jimillar RNA daga kyallen dabbobi daban-daban.
Kundin Warewa Tantanin Halitta RNA--Za'a iya samun jimlar RNA mai tsabta da inganci daga ƙwayoyin al'ada daban-daban a cikin mintuna 11.
Jumlar Shuka Kayan Keɓewar RNA--Cire RNA mai inganci da sauri daga samfuran shuka tare da ƙaramin polysaccharide da abun ciki na polyphenol.
Kit ɗin keɓewar RNA na hoto--Da sauri keɓe da tsarkake ƙwayar cuta ta RNA daga samfurori irin su plasma, serum, ruwan jiki mara-kyau da abubuwan al'adar tantanin halitta.
② Ciwon DNA na Genomic
Samfuran cirewar DNA na genomic sun haɗa da ƙasa, feces, jini, sel, kyallen dabbobi, shuke-shuke, ƙwayoyin cuta, da sauransu. Za a iya amfani da hakar DNA a cikin narkewar enzyme, ginin ɗakin karatu na DNA, PCR, shirye-shiryen antibody, Binciken haɓaka haɓakar Western blot, guntu gene, babban. -tabbataccen tsari da sauran gwaje-gwaje.
Alamar farkoKayan Keɓewar DNA
Kit ɗin Keɓewar Naman Dabbobi na DNA--Fitar da sauri da tsarkakewar DNA na genomic daga maɓuɓɓuka masu yawa, kamar kyallen dabbobi, sel, da sauransu.
Kit ɗin Midi na DNA na Jini (1-5ml)--Da sauri tsarkake high quality-genomic DNA daga anticoagulated jini (1-5ml).
Buccal Swab/FTA Katin DNA Keɓewa Kit--Da sauri tsarkake high quality-genomic DNA daga buccal swab/FTA Katin samfuran.
Kit ɗin keɓewar DNA na Shuka--Da sauri tsarkakewa da samun ingantaccen DNA na genomic daga samfuran shuka (ciki har da polysaccharides da samfuran tsirrai na polyphenol)
③ Ciwon Plasmid
Plasmid wani nau'i ne na ƙananan ƙwayoyin DNA na madauwari a cikin sel, wanda shine mai ɗaukar hoto na gama-gari don sake haɗuwa da DNA.Hanyar hakar plasmid ita ce cire RNA, ware plasmid daga kwayoyin halittar DNA, da cire furotin da sauran datti don samun plasmid mai tsafta.
Babban Plasmid Mini Kit--Da sauri tsarkake babban ingancin plasmid DNA daga ƙwayoyin cuta da suka canza don gwaje-gwajen ilimin halitta na yau da kullun kamar canji da narkewar enzyme.
④ Wasu nau'ikan hakar, hakar miRNA, da sauransu.
Kit ɗin keɓewar miRNA na dabba--A sauri da kuma yadda ya kamata cire ƙananan RNA na 20-200nt miRNA, siRNA, snRNA daga ƙwayoyin dabba da ƙwayoyin cuta daban-daban.
Ⅳ Abubuwan buƙatun don hakar acid nucleic da sakamakon tsarkakewas
① Don tabbatar da mutuncin tsarin farko na nucleic acid.
② Rage tsoma bakin sunadaran, sikari, lipids da sauran macromolecules
③ Kada a sami kaushi na halitta ko babban taro na ions karfe wanda zai iya hana enzyme a cikin samfuran nucleic acid.
④ RNA da sauran gurɓataccen acid nucleic yakamata a kawar da su yayin fitar da DNA, kuma akasin haka.
Lokacin aikawa: Nuwamba-24-2022
