Mukuita zvidzidzo zvemajini, tinowanzo sangana nemasamples eRNA asina kukwana, semuenzaniso, ekudzidza tumamota tudiki twemumuromo, kunyangwe sampuli-sero rimwe chete, uye sampuli dzekuchinja kwemajini kwakasiyana kunonyorwa pamazinga akaderera mumasero emunhu.Ehe, kune iyo COVID-19 bvunzo, kana swabs isiri munzvimbo chaiyo kana nguva isina kukwana panguva yesampling, saizi yemuenzaniso ichave yakaderera, ndosaka Komisheni yehutano nekuronga kwemhuri yakabuda mazuva maviri apfuura uye. akapasa bvunzo, uye kana iyo nucleic acid sampler isina kutora masampuli matanhatu, unogona kuzvitaura.
Kunzwa kweiyo reagent kwakakosha nekuti isu tine dambudziko iri kana iro dambudziko, saka chii chatingaite kuvandudza kunzwa kweRT-PCR?
Tisati takurukura zvingagadziriswa, ngatitaurei matambudziko maviri makuru nemamiriro achangotaura.
Chekutanga pane zvese, tinonetsekana nekurasikirwa kweRNA kana tiine mashoma mashoma emasero mumuenzaniso wedu.Kana kupatsanurwa kwechinyakare uye nzira dzekuchenesa dzakashandiswa, senge nzira yekoramu kana nucleic acid kunaya nzira, pane mukana mukuru wekuti mashoma sampuli arasikirwe.Imwe mhinduro ndeyekuwedzera mutakuri wemorekuru, senge tRNA, asi kunyangwe zvakadaro, hapana vimbiso yekuti kuedza kwedu kudzoreredza kwakanaka.
Saka ndeipi nzira iri nani?Sarudzo yakanaka yemaseru akagadzirwa kana microanatomical samples ndeye kushandisa yakananga lysis.
Pfungwa ndeyekutsemura maseru kwemaminetsi mashanu, kuburitsa iyo RNA mumhinduro, wobva wamisa kuita kwemaminetsi maviri, wobva wawedzera lysate zvakananga kune reverse transcription reaction kuitira kuti pasave neRNA inorasika, uye pakupedzisira isa inoguma cDNA zvakananga. mune chaiyo-nguva reaction.
Asi ko kana, nekuda kwenzvimbo yakaganhurirwa yekutanga kana diki diki yekutarisisa gene kutaura, isu tinogona kudzokorodza iyo RNA yese uye tisingape akakwana matemplate kuti tiwane yakanaka-chaiyo-nguva chiratidzo?
Muchiitiko ichi, danho rekutanga-amplification rinogona kubatsira zvakanyanya.
Izvi zvinotevera chirongwa chekuwedzera kunzwisiswa mushure mekudzokorora kunyorwa.Tisati tatanga, tinofanirwa kubvunza zasi kwemvura kuti ndezvipi zvinangwa zvatiri kufarira, kuitira kuti tigadzire maprimers ezvinangwa izvi zvekutanga-amplification.
Izvi zvinogona kuwanikwa nekugadzira primer yakasanganiswa inosvika zana peya dzekutanga uye kutenderera kwe10 kusvika ka14 nguva.Naizvozvo, Master Mix yakanyatso gadzirirwa ichi chinodiwa inodiwa kufanokudza cDNA yakawanikwa.
Chikonzero chekuisa huwandu hwematenderedzwa pakati pe10 ne14 ndechekuti iyi shoma nhamba yematenderedzwa inovimbisa kusarongeka pakati pezvakasiyana zvinangwa, izvo zvakakosha kune vaongorori vanoda huwandu hwehuwandu hwema molecular information.
Mushure mekutanga-amplification, tinogona kuwana huwandu hwakawanda hwecDNA, kuitira kuti kunzwisiswa kwekuona kumucheto-kumashure kunovandudzwa zvikuru, uye isu tinogona kutoderedza sampuli uye kuita akawanda-chaiyo-nguva PCR maitiro kuti abvise zvikanganiso zvingangoitika.
Nguva yekutumira: Kubvumbi-11-2023