Aratohu Pūtaiao Raumati: Ina Tūtaki te Ngaru Wera 40°C ki ngā Whakamātautau Ngota

Kua mau tonu te pāmahana teitei puta noa i te nuinga o Haina i ēnei rā tata nei. I te 24 o Hūrae, i whakaputa te Shandong Provincial Meteorological Observatory i tētahi whakatūpato pāmahana teitei kōwhai, e tohu ana i ngā pāmahana "rite ki te sauna" o te 35-37°C (111-133°F) me te 80% te haumākū mō ngā rā e whā e whai ake nei i ngā rohe o uta. Kei te tata te pāmahana i ngā wāhi pēnei i a Turpan, Xinjiang, ki te 48°C (111-133°F). Kei raro i te whakatūpato karaka a Wuhan me Xiaogan, Hubei, me te pāmahana neke atu i te 37°C i ētahi wāhi. I roto i tēnei wera nui, kei te pāngia te ao moroiti i raro i te mata o ngā pipette e ngā raruraru rerekē—ko te pumau o ngā waikawa karihi, te mahi a ngā whākōkī, me te āhua tinana o ngā matū ka whakapōturitia katoatia e te ngaru wera.

Kua rite te tango waikawa karihi ki te whakataetae ki te wā. Ina neke ake te pāmahana o waho i te 40°C, ahakoa kei te whakamahia te hauhautanga, ka neke ake te pāmahana o te tēpu mahi i te 28°C. I tēnei wā, ka pakaru ngā tauira RNA e waiho ana i waho i te puna me te ngahuru, neke atu i te rua te tere o te pirau i te puna me te ngahuru. I roto i te tango i ngā pire aukume, ka mākū te otinga parepare i te rohe nā te tere o te mimiti o te whakarewa, ā, ka ngawari te puta o ngā tioata. Ka nui te pikinga o te pai o te hopu waikawa karihi i ēnei tioata. Ka piki ake te mimiti o ngā whakarewa waro i te wā kotahi. I te 30°C, ka piki ake te nui o te mimiti o te chloroform mā te 40% ki te 25°C. I te wā e whakahaerehia ana, me whakarite kia 0.5m/s te tere o te hau i roto i te taupoki auahi, ā, me whakamahi karapu nitrile hei pupuri i te whai huatanga tiaki.

He uaua ake ngā raruraru pāmahana e pā ana ki ngā whakamātautau PCR. He tino tairongo ngā tauhohenga pēnei i te whākōkī Taq me te transcriptase whakamuri ki ngā huringa ohorere o te pāmahana. Ka taea e te haukū i runga i ngā pakitara o te ngongo i muri i te tango mai i te pouaka tio -20°C te whakaheke i te mahi whākōkī neke atu i te 15% mēnā ka uru ki te pūnaha tauhohenga. Ka taea hoki e ngā otinga dNTP te whakaatu i te hekenga kitea i muri i te 5 meneti noa iho o te whakaraeraetanga ki te pāmahana rūma (>30°C). Ka whakararuhia hoki te mahi a te taputapu e ngā pāmahana teitei. Ina neke atu i te 35°C te pāmahana taiao o te taiwhanga, ā, kāore i te rawaka te ātete wera o te taputapu PCR (<50 cm mai i te pakitara), ka taea e te rerekētanga pāmahana o roto te eke ki te 0.8°C. Ka taea e tēnei rerekētanga te whakaheke i te pai o te whakanui i te taha o te pereti 96-puna neke atu i te 40%. Me horoi tonu ngā tātari puehu (ka whakaitihia e te kohikohinga puehu te pai o te wera mā te 50%), ā, me karohia te hauhautanga tika. Hei tāpiri, i te wā e mahi ana i ngā whakamātautau PCR i te pō, karohia te whakamahi i te taputapu PCR hei "pouaka whakamātao taupua" hei penapena tauira. Ki te rongoatia i te 4°C mō te neke atu i te 2 hāora i te pāmahana, ka puta te waikura i muri i te katinga o te taupoki wera, ka waimeha te pūnaha tauhohenga, ā, ka waikura pea ngā waeine whakarewa o te taputapu.

I te mea kei te tūpono tonu ngā whakatūpato pāmahana teitei, me whakatangihia hoki e ngā taiwhanga ngota ngota te whakaoho. Me rongoa ngā tauira RNA utu nui ki roto i te pouaka tio -80°C, me te whakaurunga ki ngā wā pāmahana teitei anake. Mā te whakatuwhera i te tatau o te pouaka tio -20°C neke atu i te rima ngā wā i te rā ka nui ake te piki me te heke o te pāmahana. Me whai wāhi te iti rawa o te 50 cm o te wera ki ngā taha e rua me ngā taha o muri mō ngā taputapu whakaputa wera nui. Hei tāpiri, me whakarite anō te wā whakamātautau: 7:00-10:00 AM mō ngā mahi tairongo pāmahana pēnei i te tango RNA me te uta qPCR; 1:00-4:00 PM mō ngā mahi kore-whakamātautau pēnei i te tātari raraunga. Ka taea e tēnei rautaki te ārai i ngā tihi pāmahana teitei mai i te whakararuraru i ngā mahi nui.

He whakamātautau i te tikanga me te manawanui ngā whakamātautau ngota i te wā o te werawera. I raro i te rā raumati mutunga kore, kua tae pea te wā ki te whakatakoto i tō pipette me te tāpiri i tētahi pouaka hukapapa anō ki ō tauira kia taea ai e te taputapu te tuku i te wera nui ake. Ko tēnei whakaute ki ngā huringa pāmahana te kounga taiwhanga tino utu nui i ngā marama raumati werawera—nā te mea, i te werawera 40°C o te raumati, me whai "rohe porowhita horihori" kia tiakina pai.