Comet Assay: Njira Yomvera Yozindikira Kuwonongeka kwa DNA ndi Kukonza

Comet Assay (Single Cell Gel Electrophoresis, SCGE) ndi njira yachangu komanso yofulumira yomwe imagwiritsidwa ntchito pozindikira kuwonongeka kwa DNA ndikukonzanso m'maselo amodzi. Dzina lakuti "Comet Assay" limachokera ku mawonekedwe a comet-like mawonekedwe omwe amawoneka muzotsatira: phata la selo limapanga "mutu," pamene zidutswa zowonongeka za DNA zimasuntha, kupanga "mchira" wofanana ndi comet.

Mfundo yofunika

Mfundo ya Comet Assay imachokera ku kusamuka kwa zidutswa za DNA m'munda wamagetsi. DNA yosasunthika imakhalabe mkati mwa selo, pamene DNA yowonongeka kapena yogawanika imasamukira ku anode, kupanga "mchira" wa comet. Kutalika ndi kulimba kwa mchira kumayenderana mwachindunji ndi kuwonongeka kwa DNA.

Ndondomeko

1. Kukonzekera Maselo: Maselo oti ayesedwe amasakanizidwa ndi agarose otsika kwambiri ndipo amawayala pazithunzi za maikulosikopu kuti apange wosanjikiza wofanana.
2. Cell Lysis: Zithunzizi zimamizidwa mu njira ya lysis kuchotsa nembanemba ya cell ndi nembanemba ya nyukiliya, poyera DNA.
3. Electrophoresis: Zithunzizo zimayikidwa mu chipinda cha electrophoresis pansi pa zinthu zamchere kapena zandale. Zidutswa za DNA zowonongeka zimasamukira ku electrode yabwino mothandizidwa ndi magetsi.
4. Kudetsa: Pambuyo pa electrophoresis, zithunzizo zimadetsedwa ndi utoto wa fulorosenti (mwachitsanzo, ethidium bromide) kuti muwone DNA.
5. Microscopic Analysis: Pogwiritsa ntchito microscope ya fluorescence kapena mapulogalamu apadera, mawonekedwe a comet amawunikidwa, ndipo magawo monga kutalika kwa mchira ndi kulimba kwake amayesedwa.

Chithunzi chochokera ku biorender

Kusanthula Zambiri

Zotsatira zochokera ku Comet Assay zimawunikidwa kutengera magawo angapo:

• Utali wa Mchira: Imaimira mtunda umene DNA imasamuka, kusonyeza kukula kwa kuwonongeka kwa DNA.
• Zomwe zili mu DNA: Peresenti ya DNA yomwe imasamukira kumchira, yomwe nthawi zambiri imagwiritsidwa ntchito ngati muyeso wa kuchuluka kwa kuwonongeka kwa DNA.
• Olive Tail Moment (OTM): Zimaphatikiza kutalika kwa mchira ndi zomwe zili mu DNA kuti ipereke chidziwitso chokwanira cha kuwonongeka kwa DNA.

Mapulogalamu

1. Maphunziro a Genotoxicity: Comet Assay imagwiritsidwa ntchito kwambiri powunika zotsatira za mankhwala, mankhwala, ndi ma radiation pa DNA yama cell, ndikupangitsa kuti ikhale chida chofunikira kwambiri pakuyesa kwa genotoxicity.
2. Environmental Toxicology: Imathandiza kuwunika momwe zoipitsa chilengedwe zimakhudzira DNA ya zamoyo, kupereka chidziwitso pachitetezo cha chilengedwe.
3. Kafukufuku wa Zamankhwala ndi Zachipatala: Comet Assay imagwiritsidwa ntchito pophunzira njira zokonzera DNA, khansa, ndi matenda ena okhudzana ndi DNA. Imawunikanso momwe chithandizo cha khansa chimakhudzira ngati radiotherapy ndi chemotherapy pa DNA.
4. Sayansi ya Chakudya ndi Ulimi: Amagwiritsidwa ntchito pounika chitetezo cha mankhwala ophera tizilombo, zowonjezera zakudya, ndi zinthu zina, ndikuwunika momwe amawonongera nyama.

Ubwino wake

• Kutengeka Kwambiri: Imatha kuzindikira kuwonongeka kochepa kwa DNA.
• Ntchito Yosavuta: Njirayi ndi yowongoka, yomwe imapangitsa kuti ikhale yoyenera kuwonetsetsa kwapamwamba.
• Ntchito Yonse: Itha kugwiritsidwa ntchito pamitundu yosiyanasiyana yama cell, kuphatikiza ma cell a nyama ndi zomera.
• Quantification Mavuto: Popereka chidziwitso chokhudza kuwonongeka kwa DNA, kusanthula kachulukidwe kumadalira kwambiri mapulogalamu ndi njira zowunikira zithunzi.
• Zoyeserera: Zotsatira zimatha kutengera zinthu monga nthawi ya electrophoresis ndi pH, zomwe zimafuna kuwongolera mosamala mikhalidwe yoyesera.

Zolepheretsa

Comet Assay ndi chida chamtengo wapatali pa kafukufuku wa zamankhwala, sayansi ya chilengedwe, ndi chitukuko cha mankhwala chifukwa cha kusinthasintha kwake komanso kukhudzidwa kwakukulu pozindikira kuwonongeka ndi kukonza kwa DNA. Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology)imapereka chipinda chopingasa cha electrophoresis cha comet assay. Takulandilani kuti mulankhule nafe kuti tikambirane zaComet Assayprotocol.

Beijing Liuyi Biotechnology Co. Ltd (Liuyi Biotechnology) yakhala ikupanga zida za electrophoresis kwa zaka zopitilira 50 ndi gulu lathu laukadaulo laukadaulo ndi R&D pakati. Tili ndi mzere wodalirika komanso wathunthu wopanga kuchokera pamapangidwe mpaka kukayendera, ndi nyumba yosungiramo zinthu, komanso chithandizo chamalonda. Zogulitsa zathu zazikulu ndi Electrophoresis Cell (thanki / chipinda), Electrophoresis Power Supply, Blue LED Transilluminator, UV Transilluminator, Gel Image & Analysis System etc. Timaperekanso zida za labu monga PCR chida, vortex chosakanizira ndi centrifuge kwa labotale.

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