I-Comet Assay (I-Single Cell Gel Electrophoresis, SCGE) iyindlela ebucayi nesheshayo esetshenziswa ngokuyinhloko ukuthola ukulimala kwe-DNA nokulungisa kumaseli ngamanye. Igama elithi "Comet Assay" livela esimweni esifana nenkanyezi enomsila esivela emiphumeleni: i-nucleus yeseli yakha "ikhanda," kuyilapho izingcezu ze-DNA ezilimele zifuduka, zenze "umsila" ofana nenkanyezi enomsila.
Isimiso
Umgomo we-Comet Assay usekelwe ekufudukeni kwezingcezu ze-DNA endaweni kagesi. I-DNA engaguquki ihlala ku-nucleus yengqamuzana, kuyilapho i-DNA eyonakele noma eyizicucu ithuthela e-anode, yenze “umsila” wenkanyezi enomsila. Ubude nokuqina komsila kuhambisana ngokuqondile nezinga lomonakalo we-DNA.
Inqubo
- Ukulungiselela Iseli: Amaseli okufanele ahlolwe axutshwe ne-agarose yephoyinti elincibilikayo eliphansi futhi asatshalaliswa kumaslayidi esibonakhulu ukuze akhe ungqimba olufanayo.
- Iseli Lysis: Amaslayidi acwiliswa kusixazululo se-lysis ukuze asuse ulwelwesi lweseli kanye nolwelwesi lwenuzi, obala i-DNA.
- I-Electrophoresis: Amaslayidi abekwe egunjini le-electrophoresis ngaphansi kwezimo ze-alkaline noma ezingathathi hlangothi. Izingcezwana ze-DNA ezilimele zifudukela ku-electrode enhle ngaphansi kwethonya lenkambu kagesi.
- Ukufaka amabala: Ngemva kwe-electrophoresis, amaslayidi angcoliswa udayi we-fluorescent (isb, i-ethidium bromide) ukuze ubone ngeso lengqondo i-DNA.
- Ukuhlaziywa Kwe-Microscopic: Kusetshenziswa isibonakhulu se-fluorescence noma isofthiwe ekhethekile, ukwakheka kwenkanyezi enomsila kuyahlaziywa, futhi imingcele efana nobude nokuqina komsila kuyalinganiswa.
Isithombe esivela ku-biorender
Ukuhlaziywa Kwedatha
Imiphumela evela ku-Comet Assay iyahlolwa ngokusekelwe kumapharamitha angukhiye ambalwa:
- Ubude Bomsila: Imelela ibanga i-DNA elihamba ngayo, okubonisa izinga lomonakalo we-DNA.
- Okuqukethwe kwe-DNA yomsila: Amaphesenti e-DNA athuthela emsileni, ngokuvamile asetshenziswa njengesilinganiso somthamo womonakalo we-DNA.
- I-Olive Tail Moment (OTM): Ihlanganisa kokubili ubude bomsila nomsila okuqukethwe kwe-DNA ukuze kunikeze isilinganiso esibanzi somonakalo we-DNA.
Izinhlelo zokusebenza
- Izifundo ze-Genotoxicity: I-Comet Assay isetshenziswa kabanzi ukuhlola imiphumela yamakhemikhali, izidakamizwa, nemisebe ku-DNA yeseli, okuyenza ibe ithuluzi elibalulekile lokuhlola ubuthi be-genotoxicity.
- I-Toxicology Yezemvelo: Isiza ekuhlaziyeni umthelela wokungcola kwemvelo ku-DNA yezinto eziphilayo, inikeze imininingwane mayelana nokuphepha kwe-ecosystem.
- Ucwaningo Lwezokwelapha Nokwelapha: I-Comet Assay isetshenziswa ekutadisheni izindlela zokulungisa i-DNA, umdlavuza, nezinye izifo ezihlobene ne-DNA. Iphinde ihlole umthelela wemithi yokwelapha umdlavuza njenge-radiotherapy kanye ne-chemotherapy ku-DNA.
- Isayensi Yokudla Nezolimo: Isetshenziselwa ukuhlola ukuphepha kwezibulala-zinambuzane, izithasiselo zokudla, nezinye izinto, kanye nokutadisha imiphumela yazo enobuthi kumamodeli ezilwane.
Izinzuzo
- Ukuzwela Okuphezulu: Iyakwazi ukuthola amazinga aphansi omonakalo we-DNA.
- Ukusebenza Okulula: Indlela yokusebenza iqondile, iyenza ifaneleke ekuhlolweni kokusebenza okuphezulu.
- Uhlelo lokusebenza olubanzi: Ingasetshenziswa ezinhlotsheni ezihlukahlukene zamaseli, kuhlanganise namaseli ezilwane nezitshalo.
- Quantification Izinselele: Ngenkathi ihlinzeka ngedatha yekhwalithi ngomonakalo we-DNA, ukuhlaziya ubuningi kuncike kakhulu kuma-software nezindlela zokuhlaziya izithombe.
- Izimo Zokuhlola: Imiphumela ingathonywa izici ezifana nesikhathi se-electrophoresis ne-pH, edinga ukulawulwa ngokucophelela kwezimo zokuhlola.
Ukulinganiselwa
I-Comet Assay iyithuluzi eliyigugu ocwaningweni lwe-biomedical, isayensi yezemvelo, nokuthuthukiswa kwezidakamizwa ngenxa yokuguquguquka kwayo nokuzwela okuphezulu ekutholeni umonakalo nokulungisa i-DNA. I-Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology)inikeza igumbi le-electrophoresis elivundlile lokuhlola inkanyezi enomsila. Siyakwamukela ukuxhumana nathi ukuze sixoxe mayelanaI-Comet AssayIphrothokholi.
I-Beijing Liuyi Biotechnology Co. Ltd (i-Liuyi Biotechnology) ikhethekile ekukhiqizeni amathuluzi e-electrophoresis iminyaka engaphezu kwengu-50 neqembu lethu lezobuchwepheshe eliqeqeshiwe kanye nesikhungo se-R&D. Sinomugqa wokukhiqiza onokwethenjelwa nophelele kusukela ekuklanyweni kuya ekuhlolweni, kanye nendawo yokugcina impahla, kanye nokwesekwa kokumaketha. Imikhiqizo yethu eyinhloko i-Electrophoresis Cell (ithangi/igumbi), i-Electrophoresis Power Supply, i-Blue LED Transilluminator, i-UV Transilluminator, i-Gel Image & Analysis System njll. Siphinde sinikeze amathuluzi aselabhu njenge-PCR instrument, i-vortex mixer kanye ne-centrifuge yaselabhorethri.
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Isikhathi sokuthumela: Sep-20-2024