Kunyangwe mufundisi, mumwe mamorekuru eTemplate aizove akakwana, huwandu hwakakura kwazvo DNA vanowanzo shandiswa kune yakasarudzika PCR, semuenzaniso, kusvika 1 μg yeGenomic Mammolian DNA uye zvishoma se 1 PG yePlasmid DNA. Iyo yakajeka huwandu hunoenderana zvakanyanya pane huwandu hwemakopi eyekutevedzana kwekutevedzana, pamwe nekuoma kwayo.
Kana iyo shoma kwazvo template inoshandiswa, kuwedzera kunoenderana muhuwandu hwekugadzira maCycle zvichadiwa kuti uwane huwandu hwakakwana hwechigadzirwa. A Taq Polymerase iyo inoshandiswa kune yakawanda plR maedzo hairatidze basa rekururamisa (3'-5 'exonuclect basa); Nekudaro, zvikanganiso zviri kuitika panguva yekuwedzera hazvigone kurangwa. Iyo yepamusoro nhamba yejekisiti, iyo yakawanda yakakurumbira kukwirisa kwechigadzirwa chakakanganisika zvichave. Kana, kune rimwe divi, huwandu hwe template hwakanyanya, mukana wevanhurume unonyora kune mamwe (kwete zana muzana muzana) kutevedzana, pamwe nekuumbwa kwePrimer dimers, kuchawedzera, izvo zvichaguma nekuwedzera kwe ne-zvigadzirwa. Muzviitiko zvakawanda, DNA iri kusarudzika kubva kune tsika tsika kana kubva kune microorganisms uye yakazoshandiswa seprance steplate. Kutevera kucheneswa, kunokosha kuti uzive kushungurudzika kweDNA kuti ugone kutsanangura huwandu hwehuwandu hunodiwa kune iyo PCR setup. Nepo Agalose Gel Electrophoresis inogona kushanda kuti ipe fungidziro, nzira iyi iri kure neiyo chaiyo. UV-VIVE Spectrophotserry yakagadzwa segoridhe yegoridhe yekudzikiswa kwema acclic acids; Izvi zvakananga uye saka nyore uye nekukurumidza nzira matanho ekunyora kwemuenzaniso pa260 nm, uye kusunganidzwa kunoverengerwa nerubatsiro rwekutendeuka kwechinhu chekushandurwa.
Kana iyo DNA iyo yakaderera kwazvo, zvisinei ( Panyaya yekukatyiswa kwazvo, kuverenga kuchawedzera kuve kusiri kwekushandiswa, uye kusvibiswa kuchatungamira ku (dzimwe nguva kwakakura) kuwedzeredza kukosha chaiko. Mune ino kesi, huwandu uchishandisa fluorescence inogona kuburitsa imwe nzira. Iyi nzira inoenderana nekushandiswa kweiyo fluorescent dhayi iyo inosungira zvakananga kuDSDN chete iyo yakaomarara yekukanda acid uye dhayi inofara neyakajeka yemwenje yakakwira wavelength. Pano, kusimba kweiyo fluorescent chiratidzo kuenderana nehuwandu hweDNA, uye nekuona iyo yekuvhara iyo inoongororwa maererano neyakajairwa curve. Zvakanakira nzira iyi zorora pane chokwadi chechisungo, icho chisingabvumirani nezvekunze kwenyika kwakaunzwa nekusvibiswa, pamwe nekukwanisa kwekugona kuona zvakanyanya kuderera kweDNA. Kukodzera kweimwe nzira kunoenderana zvakanyanya pamhirizhonga uye kuchena; Muzviitiko zvakawanda zvinogona kunge zvichitombove zano rekushandisa nzira mbiri dziri kufanana.
Kutumira Nguva: Nov-30-2022