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He Tikanga Āhua-Matū Aukume e Whakatau Pai ana i ngā Wero o te Tangohanga DNA Wai Taiao
I roto i ngā mara pēnei i te rangahau moroiti taiao me te aroturuki i te parahanga wai, ko te tango i te DNA ira kounga teitei he mea nui mō ngā tono o muri mai tae atu ki te PCR/qPCR me te raupapatanga o te whakatupuranga e whai ake nei (NGS). Heoi, he tino uaua ngā tauira wai taiao, kei roto ngā hapori moroiti kanorau, ngā momo uaua ki te wetewete pēnei i te huakita Gram-pai, me ngā wero roa e pā ana ki ngā tikanga tango tuku iho—pēnei i te whakamahinga o ngā matū paitini me ngā tikanga uaua—kua whakararuraru tonu i ngā kairangahau.
Ināianei, ka whakaurua mai e Bigfish Sequencing te BFMP24R Magnetic Bead-Based Environmental Water Genomic DNA Extraction and Purification Kit, e whakarato ana i tētahi otinga whānui ki ēnei wero mā te hangarau auaha me te hoahoa ngāwari ki te whakamahi.
Tirohanga Whānui o te Hua
Ko tēnei kete he mea hanga i runga i tētahi pūnaha ārai kua arotauhia me ngā pīrere aukume nano mahi-teitei. Ka herea motuhaketia te ira ira ki ngā rōpū mahi i runga i te mata o te pīrere, ā, ka wehea i raro i tētahi papa aukume o waho. Whai muri i ngā mahi horoi mārie maha hei tango i ngā pūmua, ngā tote, me ētahi atu poke, ka oti te whakakore i te ira ira tino-parakore.
He mea hanga motuhake mō ngā tauira wai taiao, he pai te tango a te kete i te DNA huakita i kohia i runga i ngā kiriuhi tātari, tae atu ki ngā huakita Gram-kino me ngā huakita Gram-pai (tae atu ki te 2 × 10⁹ pūtau huakita mō ia kiriuhi tātari kotahi). He hototahi ki ngā pūnaha tango waikawa karihi aunoa katoa mō te tukatuka tere. He rite tonu te kounga o te DNA i tangohia, ā, ka taea te whakamahi tika mō te PCR/qPCR, NGS, me ētahi atu tono o muri mai.
Ngā Āhuatanga o te Hua
1. Te Kaha Tango Huakita Whānui
He whai hua te tango i ngā huakita Gram-kino me ngā huakita Gram-pai mai i ngā tauira wai, e kapi ana i ngā hapori moroiti e kitea nuitia ana i ngā taiao wai māori me te moana, ā, e tutuki ana hoki i ngā whakaritenga tātaritanga kanorau.
2. Teitei o te Parakore me te Hua Nui
Ka tukuna te DNA he tino parakore, kāore he poke aukati, ā, he hua pumau e tika ana mō ngā tono ngota ngota tika i raro iho.
3. Hototahi Aunoa me te Tino Whai Hua
He tino hototahi ki ngā pūnaha tango waikawa karihi aunoa a Bigfish, e tautoko ana i te tukatuka wā kotahi o ngā tauira 32, 96 rānei, e whakaiti ana i te wā tukatuka me te whakapai ake i te whai huatanga o te taiwhanga.
4. Mahi Haumaru, Ātaahua hoki
Kāore e hiahiatia he matū paitini pēnei i te phenol, te chloroform rānei, hei whakaiti i ngā mōrearea haumaru taiwhanga. Kua takaihia ngā matū matua ki roto i ngā pereti 96-pū, hei whakaiti i ngā hapa o te tango paipa ā-ringa, hei whakahaere hoki i ngā rerenga mahi.
Ngā Pūoro Hototahi
Ika Nui BFEX-16E
BFEX-32
BFEX-32E
BFEX-96E
Ngā Hua Whakamātautau
I tātarihia he tauira wai awa 600 mL mā roto i tētahi kiriuhi, ā, i tangohia te DNA mā te whakamahi i te kete tango me te purenga DNA ira taiao wai ā-aukati Bigfish me te taputapu hototahi. I tātarihia te DNA i tangohia mā te electrophoresis agarose gel.
M: Tohu1, 2: Ngā tauira wai awa
Ngā Whakatakotoranga Hua
Wā tuku: Hakihea-18-2025
Ngā tautuhinga tūmataiti