Kodi centrifuge imasiyanitsa DNA ndi mapuloteni?

Kodi Centrifuge imasiyanitsa DNA ndi Mapuloteni?

Chiyambi

Kugawaniza maselo ndi njira yofunika kwambiri yomwe imagwiritsidwa ntchito kwambiri mu kafukufuku wa sayansi ndi ma laboratories azachipatala pogawaniza zigawo zosiyanasiyana za chitsanzo kutengera kuchuluka kwawo. Njira imodzi yomwe imagwiritsidwa ntchito kwambiri pa njirayi imaphatikizapo kupatula DNA ndi mapuloteni kuchokera ku chitsanzo chachilengedwe. M'nkhaniyi, tifufuza ngati centrifuge imatha kulekanitsa DNA ndi mapuloteni bwino ndikufufuza njira zosiyanasiyana zomwe zimagwiritsidwa ntchito kuti pakhale kulekanitsa koyenera.

Kumvetsetsa Centrifugation

Musanafufuze mfundo zenizeni zolekanitsa DNA ndi mapuloteni pogwiritsa ntchito centrifuge, ndikofunikira kumvetsetsa mfundo zazikulu zoyendetsera centrifuge. Centrifuge imagwira ntchito motsatira mfundo yoyendetsera matope ndipo imagwiritsa ntchito kupota kwachangu kuti ipange mphamvu yamphamvu yoyendetsera centrifuge. Mphamvu imeneyi imapangitsa kuti tinthu tating'onoting'ono mu chitsanzo tisiyane kutengera kuchuluka kwawo, ndipo tinthu tating'onoting'ono timasunthira pansi pa chubu cha chitsanzo. Chifukwa chake, tinthu tating'onoting'ono timakhala pamwamba.

Vuto Lolekanitsa DNA ndi Mapuloteni

Ponena za kulekanitsa DNA ndi mapuloteni, zimakhala zovuta chifukwa mamolekyu onse awiri ali ndi kulemera kofanana kwa mamolekyu. Popanda njira yothandiza yolekanitsa, zimakhala zovuta kupeza zitsanzo za DNA yeniyeni kapena mapuloteni. Komabe, centrifugation ingathandize pakulekanitsa, koma sizingakhale zokwanira zokha.

1. Kusiyanitsa kwa Differential Centrifugation: Kupatukana Koyamba

2. Kuchepetsa mphamvu zamagetsi: Kulekanitsa kowonjezereka

3. Kuchepetsa Mphamvu ya Maginito: Kukonza Njira

4. Kufunika kwa Kusankha Buffer

5. Kuchepetsa mphamvu ya centrifugation poyerekeza ndi njira zina zolekanitsira

Kusiyana kwa Centrifugation: Kupatukana Koyamba

Kusiyanitsa kwa ma centrifugation ndi gawo loyamba pakulekanitsa DNA ndi mapuloteni. Njirayi imagwiritsa ntchito liwiro losiyana la ma centrifugation kuti ikwaniritse kulekanitsa kwakukulu kwa zigawo za maselo. Pa nthawiyi, maselo ogwirizana kapena zitsanzo za minofu zimayendetsedwa ndi centrifugation yothamanga kwambiri, nthawi zambiri pafupifupi 1000-5000 rpm (rpm). Kuzungulira kothamanga kumeneku kumathandiza kulekanitsa ma nuclei ndi maselo osatha, zomwe zimapangitsa kuti pakhale kugawikana kwa crude. Komabe, sikupereka kulekanitsa kwathunthu kwa DNA ndi mapuloteni.

Kuchepetsa mphamvu zamagetsi: Kulekanitsa Kwambiri

Kuti athetse zofooka za differential centrifugation, ultracentrifugation imayamba. Ultracentrifuges ndi ma centrifuge othamanga kwambiri omwe amatha kufika pa liwiro la 100,000 rpm. Mwa kuyika gawo lopanda kanthu lomwe lapezeka kuchokera ku centrifugation yoyambirira yothamanga pang'ono kupita ku ultracentrifugation, kusiyana kwakukulu pakati pa DNA ndi mapuloteni kungatheke. Mamolekyu olemera a DNA amagwera pansi pa chubu, ndikupanga pellet yooneka, pomwe mapuloteni amakhalabe mu supernatant.

Kuchuluka kwa Gradient Centrifugation: Kukonza Njira

Kuti muwongolere kulekanitsa kwa DNA ndi mapuloteni, kugwiritsa ntchito njira yochepetsera kuchuluka kwa ma protein pogwiritsa ntchito density gradient centrifugation. Mu njira iyi, njira yochepetsera kuchuluka kwa ma protein, monga sucrose kapena cesium chloride, imakonzedwa. Njirayi imakhala ndi ma densities osiyanasiyana, omwe amawonjezeka kuchokera pamwamba mpaka pansi pa chubu cha centrifuge. Gawo lopanda kanthu lomwe lapezeka kuchokera pa sitepe yapitayi limayikidwa mosamala pamwamba pa density gradient ndikuyikidwanso ku ultracentrifugation kachiwiri. DNA ndi mapuloteni zimalekanitsidwa kutengera ma densities awo ndikupanga mikanda yosiyana mkati mwa gradient. Kugawa kwa gradient kumalola kuti DNA yeniyeni ndi mapuloteni zibwezeretsedwe.

Kufunika kwa Kusankha Buffer

Buffer yomwe imagwiritsidwa ntchito panthawi ya centrifugation imagwira ntchito yofunika kwambiri pakupangitsa kulekanitsidwa bwino. Imakhudza kukhazikika ndi umphumphu wa mamolekyu omwe akulekanitsidwa. Pa DNA, buffer ya Tris-EDTA yokhala ndi pH ya 8.0 imagwiritsidwa ntchito kwambiri chifukwa cha kuthekera kwake kusunga kukhazikika kwa DNA. Kumbali ina, mapuloteni amafuna ma buffer okhala ndi pH yeniyeni, nthawi zambiri pafupifupi 7.0, kuti apewe kuwonongeka kwa chilengedwe ndikusunga kapangidwe kawo. Kusankha buffer yoyenera kumatsimikizira kulekanitsidwa koyenera kwa DNA ndi mapuloteni popanda kuwononga ubwino wawo.

Kuchepetsa Kupanikizika Mosiyana ndi Njira Zina Zopatukana

Ngakhale kuti njira yoyezera DNA ndi njira yomwe imagwiritsidwa ntchito kwambiri polekanitsa DNA ndi mapuloteni, ndikofunikira kuvomereza kuti mwina si njira yokhayo yomwe imagwiritsidwa ntchito. Njira zina, monga chromatography, electrophoresis, ndi precipitation, zitha kuwonjezera njira yoyezera DNA kuti ipeze kulekanitsa kolondola kwambiri. Njira iliyonse ili ndi ubwino ndi zofooka zake, ndipo ofufuza nthawi zambiri amaphatikiza njira zingapo kuti apeze kuyera komwe kumafunikira komanso kubwezeretsa DNA ndi mapuloteni.

Mapeto

Pomaliza, ngakhale kuti centrifuge ndi yothandiza kwambiri polekanitsa DNA ndi mapuloteni, imagwira ntchito bwino kwambiri mogwirizana ndi njira zina. Kusiyanitsa kwa centrifugation, ultracentrifugation, ndi density gradient centrifugation ndi njira zingapo zolekanitsira zomwe zimathandiza kupeza zitsanzo zoyera. Kuphatikiza apo, kusankha ma buffer oyenera ndi kuganizira njira zina zolekanitsira kumawonjezeranso magwiridwe antchito onse a kulekanitsa DNA ndi mapuloteni.