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E pili ana i ka RNA extraction

He kanakolu mau makahiki i hala aku nei, no ka unuhi ʻana o RNA, ʻo ka mea hiki iā mākou ke noʻonoʻo pehea e loaʻa ai ka RNA piha, akā naʻe, ʻaʻohe koi no ka wikiwiki o ka unuhi ʻana (The first-generation Techology the Trizol method born).Me ka hoʻonui hou ʻana o ka ʻenehana unuhi RNA, ʻaʻole i kaupalena ʻia ka mea hiki iā mākou ke hana i ka loaʻa ʻana o nā hopena.I ka wā e ʻimi ai i ka unuhi ʻana o nā reagents i hoʻokumu ʻia ma Trizol, ua pilikia nā mea hoʻokolohua e kāna mau hana paʻakikī;ʻoiai ma hope o ka nui o nā loiloi a me nā hana hou, ua lōʻihi ka manawa.I ka manawa like, pono e hoʻohana ʻia nā reagents organik e like me ka phenol a me ka chloroform i ke kaʻina hana ʻokiʻoki, kahi e hoʻoweliweli pū ai i ka palekana o nā limahana kūpono i kekahi ʻano.

2Pono, kūpono a palekana

ʻO ka loaʻa ʻana o kahi huahana unuhi RNA kūpono, maʻalahi a palekana i lilo i pahuhopu mua o nā mea hoʻokolohua.No laila, ua hoʻokumu ʻia ke ʻano hana adsorption Silica Spin Column no ka ʻohi RNA.ʻO ka papa inoaʻekolu o RO Foregerane RNN . ^.

Papa hoʻohālikelike kaʻina hana

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Palapala hoʻohālikelike hopena hoʻokolohua

ʻO ka pahu hoʻokaʻawale RNA holoʻokoʻa Foregene he kiʻekiʻe RNA extraction yield, maʻemaʻe maikaʻi, a ʻoi aku ka maikaʻi o ka hopena o ka miRNA ma mua o nā pahu hōʻailona ʻē aʻe.

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Electrophoresis o 50 mg o ka ʻiole ate ka huina RNA i unuhi ʻia e nā ʻano hana like ʻole
ʻO ka hanauna mua (1: Trizol)
ʻO ka lua o ka hanauna (2: RNA extraction kit mai ka hui A)
ʻO ke kolu o ka hanauna (3: RNA extraction kits mai ka hui A, 4: Foregene:RNA Isolation kits)

Kiʻi kiʻi qPCR (kiʻi kipi hoʻonui)

Pono e hoʻoneʻe maikaʻi ʻia ka DNA Genomic i ka wā o ka unuhi ʻana o RNA, i ʻole e loaʻa i nā hopena hiki ʻole ke hoʻopaʻa ʻia i nā hoʻokolohua o lalo (ke kumu CT mua, nā hopena maikaʻi ʻole).Hiki iā Foregene ke hoʻopau pono i ka DNA genomic me ka ʻole o ka hoʻohui ʻana o DNase, e mālama ana i ka manawa he nui, ʻoiai e pale ana i ka hoʻohaʻahaʻa ʻia e ka exogenous RNase contamination.
ʻO ka hanauna mua (hōʻike ka ʻōmaʻomaʻo iā TRlzol-, hōʻike ka ʻulaʻula iā TRlzol+)

5ʻO ka lua o ka hanauna (hōʻike ʻo Blue i ka hui A RNA pahu pahu pahu -, ʻulaʻula e hōʻike ana i ka hui A RNA pahu pahu +)

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ʻO ke kolu o ka hanauna (hōʻike ka polū i ka hui A RNA extraction kits me ka ʻōmaʻomaʻo e hōʻike ana i nā kits Foregene RNA Isolation kits)

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(E hoʻomaopopo: "_" ma ka pakuhi qPCR e hōʻike ana ʻaʻole i hoʻohui ʻia ʻo DNase, a ʻaʻole i wehe ʻia ka haumia o ka DNA; "+" e hōʻike ana ua hoʻohui ʻia ka DNase, a ua wehe ʻia ka haumia o DNA)

Hoʻopiha piha i ka RNA i loko o 11min

ʻAʻole pono no nā makahiki o ka hoʻonui ʻana i ka ʻike nucleic acid, Foregene high-efficiency total RNA isolation kits series huahana hiki iā ʻoe ke loaʻa i ka RNA kiʻekiʻe a me ka maʻemaʻe kiʻekiʻe i loko o 11 mau minuke e hoʻokō i nā pono hoʻokolohua like ʻole.

9A hiki i kēia manawa, ua ʻike ʻia nā huahana foregene total RNA isolaton kits series e ka nui o nā mea kūʻai aku a ua hāʻawi i nā palapala kiʻekiʻe kiʻekiʻe.ʻO ka hoʻohana maoli ʻana o ka mea kūʻai aku i ka pakuhi electrophoresis manaʻo a me ka quantitative PCR amplification curve ma hope o ka transcription reverse e hōʻike ana ʻo ka RNA i unuhi ʻia e ka hoʻohana ʻana i nā huahana Foregene ʻaʻole kiʻekiʻe wale nō i ka noʻonoʻo ʻana, akā i ka pololei maikaʻi.ʻO ka ʻenehana huahana wale nō i hoʻāʻo ʻia e ka mākeke hiki ke makemake ʻia e kēlā me kēia.

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Kelepona Huina RNA Isolation Kit

ʻO ka hua RNA kiʻekiʻe

ʻO ka wikiwiki: hoʻopau i ka hoʻokaʻawale RNA i 11 mau minuke

Palekana: ʻAʻohe mea kemika Organic i hoʻohui ʻia

ʻO kekahi mau huaʻōlelo kiʻekiʻe:

1. Yuan Fang, Zezhong Liu, Yang Qiu, et al.ʻO ka pale ʻana i ka mea hoʻomake viral o nā protein RNAi e nā peptides mea hoʻolālā e pale ai mai ka maʻi enteroviral i vivo, Immunity, 54(10), 2021: 2231-2244.e6.(IF:31.745,Kiko Hoʻokaʻawale RNA Viral,Piki No.RE-02011)

https://www.sciencedirect.com/science/article/pii/S1074761321003617

2. Ren, Y., Wang, A., Wu, D. et al.ʻO ka pāpālua ʻana o ka pale ʻana a me ka apoptosis e ke kanaka cytomegalovirus protein UL37x1 hiki ke hoʻopili hou i ka maʻi virus.Nat Microbiol 7, 1041–1053 (2022).(IF:30.964,Kelepona Huina RNA Isolation Kit,Piki No.RE-03111)

https://doi.org/10.1038/s41564-022-01136-6


Ka manawa hoʻouna: Iulai-27-2022