Ka wehea e te centrifuge te DNA mai i ngā pūmua
Ka Wehea e te Centrifuge te DNA mai i ngā Pūmua?
Kupu Whakataki
He tikanga nui te whakamahi i te pūwero hei wehewehe i ngā wāhanga o tētahi tauira i runga i te mātotorutanga. Ko tētahi whakamahinga noa o tēnei tikanga ko te wehewehe i te DNA me ngā pūmua mai i tētahi tauira koiora. I roto i tēnei tuhinga, ka tūhuratia e mātou mēnā ka taea e te pūwero te wehewehe pai i te DNA mai i ngā pūmua, me te ruku hohonu ki ngā tikanga rerekē e whakamahia ana hei whakatutuki i te wehenga whai hua.
Te Mārama ki te Whakapūngao
I mua i te ruku hohonu ki ngā āhuatanga motuhake o te wehewehe i te DNA mai i ngā pūmua mā te whakamahi i te centrifuge, he mea nui kia mārama ki ngā mātāpono taketake o te centrifugation. Ka mahi te centrifuge i runga i te mātāpono o te whakatō, ā, ka whakamahia te hurihuri tere-teitei hei hanga i te kaha centrifuge kaha. Mā tēnei kaha ka wehe ngā matūriki i roto i te tauira i runga i tō rātou kiato, me ngā matūriki kiato e heke ana ki raro o te ngongo tauira. Nō reira, ko ngā matūriki māmā ake ka noho tonu ki runga.
Te Wero o te Wehewehe i te DNA me ngā Pūmua
Ina tae ki te wehewehe i te DNA me ngā pūmua, he uaua tēnei nā te mea he ōrite te taumaha o ngā ngota ngota e rua. Ki te kore he tikanga wehewehe whai hua, he uaua ki te tiki tauira DNA, pūmua rānei parakore. Heoi, ka taea e te centrifugation te āwhina i te tukanga wehewehe, engari kāore pea e ranea ko ia anake.
1. Te Whakapūtanga Rerekētanga: Te Wehenga Tuatahi
2. Ultracentrifugation: Wehenga Whakarei Ake
3. Te Whakapūtanga o te Rerekētanga Kiato: Te Whakapaipai i te Tukanga
4. Te Hiranga o te Kōwhiringa Pūwero
5. Te Whakapūngao me ētahi atu Tikanga Wehewehe
Te Whakapūtanga Rerekētanga: Te Wehenga Tuatahi
Ko te whakapūtanga rerekē te taahiraa tuatahi ki te wehewehe i te DNA mai i ngā pūmua. Ka whakamahia e tēnei tikanga ngā tere whakapūtanga rerekē hei whakatutuki i te wehenga whānui o ngā wāhanga pūtau. I roto i tēnei tukanga, ka tukuna ngā pūtau kua whakakotahitia, ngā tauira kiko rānei ki te whakapūtanga tere-iti, tata ki te 1000-5000 hurihanga ia meneti (rpm). Mā tēnei hurihanga tere-iti ka taea te wehenga o ngā karihi me ngā pūtau kāore i pakaru, ka hua ake he wehenga mata. Heoi, kāore e whakarato i te wehenga katoa o te DNA me ngā pūmua.
Te Whakapūngao-ā-Ultra: Te Wehenga Whakarei Ake
Hei whakakore i ngā herenga o te centrifugation rerekētanga, ka uru mai te ultracentrifugation. Ko ngā ultracentrifuges he centrifuges tere-teitei ka taea te eke ki te tere tae atu ki te 100,000 rpm. Mā te tuku i te wāhanga mata i whiwhi mai i te centrifugation tere-iti tuatahi ki te ultracentrifugation, ka taea te whakatutuki i tētahi taumata wehenga teitei ake i waenga i te DNA me ngā pūmua. Ka totohu ngā ngota DNA taumaha ki raro o te ngongo, ka hanga he pirepire e kitea ana, ko ngā pūmua kei roto tonu i te waikeri.
Te Whakapūtanga o te Rerekētanga Matotoru: Te Whakapaipai i te Tukanga
Hei whakapai ake i te wehenga o te DNA me ngā pūmua, ka whakamahia te centrifugation mātotoru. I roto i tēnei tikanga, ka whakaritea he reo mātotoru, pērā i te sucrose, te cesium chloride rānei. He whānuitanga o ngā mātotoru kei roto i tēnei reo, e piki haere ana mai i runga ki raro o te ngongo centrifuge. Ko te wāhanga mata i whiwhi mai i te taahiraa o mua ka ata whakatakotoria ki runga i te mātotoru mātotoru, ā, ka tukuna anō ki te ultracentrifugation. Ka wehea te DNA me ngā pūmua i runga i ō rātou mātotoru, ā, ka hanga he roopu motuhake i roto i te mātotoru. Mā te wehewehenga o te mātotoru ka taea te whakahoki mai i ngā tauira DNA me ngā pūmua parakore.
Te Hiranga o te Kōwhiringa Pūwero
He mea nui te mahi a te pūhui e whakamahia ana i te wā o te centrifugation ki te whakatutuki i te wehenga whai hua. Ka pā ki te pumau me te āhua o ngā ngota e wehea ana. Mō te DNA, he mea whakamahi noa te pūhui Tris-EDTA me te pH o te 8.0 nā te mea ka taea e ia te pupuri i te pumau o te DNA. I tētahi atu taha, me whai pūhui ngā pūmua me ngā taumata pH motuhake, tata ki te 7.0, hei ārai i te denaturation me te pupuri i tō rātou hanganga taketake. Mā te whiriwhiri i te pūhui tika ka tika te wehenga o te DNA me ngā pūmua me te kore e whakararu i tō rātou kounga.
Te Whakapūngao me ētahi atu Tikanga Wehewehe
Ahakoa he tikanga whānui te whakamahi i te centrifugation hei wehewehe i te DNA mai i ngā pūmua, he mea nui kia mōhio ehara pea koinei anake te tikanga e whakamahia ana. Ka taea e ētahi atu tikanga, pērā i te chromatography, te electrophoresis, me te precipitation, te tāpiri atu ki te centrifugation hei whakatutuki i tētahi wehenga pai ake. He painga me ngā herenga anō tō ia tikanga, ā, he maha ngā wā ka whakakotahi ngā kairangahau i ngā tikanga maha hei whiwhi i te parakore e hiahiatia ana me te whakaora i te DNA me ngā pūmua.
Whakamutunga
Hei whakamutunga, ahakoa he tino whai hua te centrifuge mō te wehewehe i te DNA mai i ngā pūmua, he pai ake te mahi tahi me ētahi atu tikanga. Ko te centrifugation rerekē, te ultracentrifugation, me te centrifugation gradient density e hanga ana i tētahi raupapa o ngā mahi wehewehe e āwhina ana ki te whiwhi tauira parakore. Hei tāpiri, ko te whiriwhiri i ngā arai tika me te whakaaro ki ētahi atu tikanga wehewehe ka whakarei ake i te whai huatanga whānui o te wehenga DNA-pūmua.
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