Nucleic Acid Extraction po'o le Pusa Fa'amamāina
Nucleic Acid Extraction Or Fa'amamāina Kitpe teuina ile -20℃. O le faʻataʻitaʻiga e tatau ona feaveaʻi e faʻaaoga ai le 0℃ curling.
Introduction
O le Nucleic Acid Extraction poʻo le Purification Kit (Magnetic Beads Method) ua mamanuina mo le faʻamamaina otometi o le RNA ma le DNA mai vai o le tino (e pei o swabs, plasma, serum) e faʻaaoga ai mea faʻapipiʻi otometi nucleic acid. O tekonolosi maneta e maua ai le DNA/RNA maualuga e talafeagai mo le fa'aoga sa'o i fa'aoga i lalo e pei o le fa'ateleina po'o isi gaioiga fa'a-enzymatic.
Application Range
O le toto atoa, plasma, serum ma isi fa'ata'ita'iga a le tino na fa'amama sa'o ma fa'amama. O le nucleic acid na fa'asa'olotoina na fa'apipi'iina e le super paramagnetic nanometer maneta lopa. Ona aveese lea o le polotini, ion masima inorganic ma mea eleelea e ala i le fufuluina o vaifofo. Mulimuli ane, o le nucleic acid na faʻafefe i le eluent e maua ai se vaifofo mama nucleic acid.
Kit Contents
| Pusi. Leai. | YXN-VIRAL01-32A-BR | Vaega | ||
| -50A | - 100A | |||
| Tele | 32Te | 50Su'ega | 100Su'ega | |
| Lysis Pupa | 96 lelei ed Papatusi 2 fasi | 25ml | 50ml | Surfactant ma Tris |
| Fufulu Fa'amau I | ★15ml | ★30ml | Fofo maualuga masima | |
| Fufulu Puipui Il | ★6ml*2 | ★12ml*2 | Fofo maualalo masima | |
| Elution Buffer | 10ml | 20ml | Fofo maualalo masima | |
| MagaBio Reagent | 1.0ml | 2.0ml | Maliega maneta | |
| Tusitaulima(=YXN-VIRAL01-32A-BR) | 1 | 1 | 1 | |
| Nmea:MoYXN-VIRAL01-32A-BR-50A,fa'aopoopo le 15mL Absolute ethanol ile ★15mL Wash Buffer I a'o le'i fa'aogaina; fa'aopoopo le 24mL Ethanol atoa ile ★6mL Fufulu fa'apolo Il a'o le'i fa'aoga. | ||||
| MoYXN-VIRAL01-32A-BR-100A, fa'aopoopo le 30mL Absolute ethanol i le ★30mL Wash Buffer I a'o le'i fa'aogaina; fa'aopopo le 48mL Absolute ethanol i le ★12mL Fufulu pa'u Il a'o le'i fa'aogaina.【Reagents e saunia e le tagata fa'aoga】Fa'amolemole saunia le ethanol (vaega su'esu'e) e oe lava. | ||||
Stmoli Conditions
A taunuu le pusa, e mafai ona teuina vaega o pusa i le vevela o le potu (15 - 25°C). E mautu le reagents mo le tasi le tausaga mai le aso o le gaosiga.
Sample Requirements
1. Faʻataʻitaʻiga talafeagai: Swabs, plasma, serum ma toto atoa ma isi.
2. Faʻataʻitaʻiga teuina ma felauaiga: E tatau ona faʻataʻitaʻiina vave le faʻataʻitaʻiga
Materials ma Devices Required but Not Provided
1. Totigilima e leai se pauta fa'aaoga
2. pusa Biohazard
3. Penitala po'o le per
Procedure
O lo'o fa'aaogaina e le vaega o lo'o mulimuli mai le fa'ata'ita'iga fa'ata'ita'iga o le fa'a'a'a'a'ese ga'o e fai ma fa'ata'ita'iga e fa'amatala fa'apu'upu'u le fa'agaioiga o le su'ega fa'ameamea i luga ole meafaigāluega fa'ameamea ole su'eina o le acid nucleic.Bioer NPA-32PpeSMTUSI 32. Mo isi ituaiga fa'ata'ita'iga, fa'amolemole tagai ile tusi fa'aoga. E mafai foi ona fa'agaioia e tagata fa'atau e tusa ai ma fa'ata'ita'iga mauaina:
1. Reagent Sauniuniga
a. MoYXN-VB03-32A-50A ma YXN-VB03-32A-100A
Faaopopo le 500uL Lysis Buffer i le koluma 1 ma le 7 o le 2.2mL 96-deep-well plate, 500uL Wash Buffer I i le koluma 2 ma le 8, 500uL Wash Buffer II i le koluma 3, 4 ma le 9,10; 70uL Elution Buffer i le koluma 5 ma le 11, 180uL vai mama ma le 20uL MagaBio Reagent i le koluma 6 ma le 12 (e tatau ona fa'afefiloi lelei ia lopa mageta a'o le'i fa'aogaina),
e. MoYXN-VB03-32A
Tu'u le 96 mea fa'apipi'i lelei i le vevela o le potu. Lulu le ipu 96-vai i luga i lalo mo le tolu taimi, ma sasae ese le taga pepa iila. Centrifuge le reagent ua uma ona teuina mo ni nai sekone (poo le sasau i le lima mo ni nai taimi) e aloese ai mai le pipii atu i le puipui o paipa. Ta'e ese le ata alumini pepa o le ipu 96-vaivai ma faailoa le itu o le ipu (lopa maneta i le koluma #6 ma le #12),
2.Fa'ata'ita'iga Ave'esea
1. Fa'aopoopo le 300uL fa'ata'ita'iga ile 96 va'alele ipu koluma #1 ma le #7, fa'amolemole 'alo ese mai fa'ama'i pipisi,
2. Tuu le ipu vaieli loloto 96 i le meafaigaluega, faʻapipiʻi le 8-strip tips i luga o le meafaigaluega,
3. Fa'agasolo le polokalame e tusa ai ma fa'atonuga nei,
4. A maeʻa le faʻamamaina otometi, faʻafeiloaʻi le elution Buffer i koluma 5 ma le 11 i se faʻamau mama 0.5mL centrifuge tube; afai e le faʻaaogaina vave, faʻamolemole teu i le-20 °C.
Performance Characteristics
1. O le oloa maua mai e iloa e le maualuga-maaleale HBV DNA suʻesuʻega reagent e oʻo atu i se lagona o le 10 IU / mL. O le oloa maua mai e iloa e le maualuga-maaleale HCV RNA detection reagent e oʻo atu i se lagona o le 50 IU/mL.
2. Filifili 4 faʻataʻitaʻiga (serum / plasma sample, nasopharyngeal swab sample, cervical exfoliated cell sample), o faʻataʻitaʻiga taʻitasi e faʻafefeteina 10 taimi ma 3 gradients (e aofia ai le faʻataʻitaʻiga muamua o le aofaʻi o 4 concentrations), faʻaaoga reagents agavaa ma suʻega suʻega e iloa ai totonu. gene fa'asinomaga e tusa ai ma fa'atonuga o oloa, ma o le tau o le Ct o vaega ta'itasi e ese i lalo ifo o le 1.
| Step | Ia Nofoaga | Program Igoa | Waiting Time(min:SS) | Mixing Time(mi totonu:SS) | Magnet Time(min:SS) | Adsorption | Spi | Volume tatu(μL) | Temperature |
| 1 | 1 | Lytuafafine | 0:00 | 2:00 | 0:00 | F | 700 | 80 | |
| 2 | 6 | Btaliga | 0:00 | 0:15 | 0:15 | √ | F | 200 | |
| 3 | 1 | Bind | 0:00 | 3:00 | 0:45 | √ | F | 700 | |
| 4 | 2 | Wlefulefu1 | 0:00 | 0:30 | 0:30 | √ | F | 500 | |
| 5 | 3 | Wlefulefu2 | 0:00 | 0:30 | 0:30 | √ | F | 500 | |
| 6 | 4 | Wlefulefu3 | 0:00 | 0:30 | 0:30 | √ | F | 500 | |
| 7 | 5 | Elution | 2:00 | 2:30 | 0:30 | F | 70 | 80 | |
| 8 | 6 | lafoai | 0:00 | 0:15 | 0:00 | F | 200 |
Safety
1. GENERAL SAOGALEMU.
O le fa'aogaina o lenei oloa i se faiga e le o fa'amaoti mai i totonu o fa'amaumauga a le tagata fa'aoga e ono i'u ai i manu'a po'o le fa'aleagaina o le meafaigaluega po'o le masini. Ia mautinoa o so'o se tasi o lo'o fa'aaogaina lenei oloa na maua fa'atonuga i faiga masani mo le saogalemu mo fale su'esu'e ma fa'amatalaga saogalemu o lo'o tu'uina atu i lenei pepa.
1.1 Aʻo leʻi faʻaaogaina se meafaigaluega poʻo se masini, faitau ma malamalama i faʻamatalaga saogalemu o loʻo tuʻuina atu i totonu o faʻamaumauga a le tagata faʻaoga na tuʻuina atu e le gaosiga o le meafaigaluega poʻo le masini.
1.2 A'o le'i tagofia vaila'au, faitau ma malamalama i Pepa Fa'amaumauga uma o le Saogalemu (SDSs) talafeagai ma fa'aoga mea e puipui ai le tagata lava ia (totini lima, ofu talaloa, puipuiga mata, ma isi). Mo le mauaina o SDSs, tagai i le vaega “Documentation and Support” i lenei pepa.
2. Kemisi saogalemu
FA'ATAUINA FA'AVAE FA'AVAE. Ina ia fa'aitiitia lamatiaga, fa'amautinoa e faitau ma fa'atino e tagata su'esu'e ta'iala lautele mo le fa'aogaina o vaila'au, teuina, ma otaota o lo'o tu'uina atu i lalo, ma fa'afeso'ota'i le SDS talafeagai mo puipuiga ma fa'atonuga fa'apitoa: Faitau ma malamalama i Pepa Fa'amaumauga o le Saogalemu (SDS) o lo'o saunia e le vaila'au. gaosi oloa ae e te le'i teuina, tago, pe galue i so'o se vaila'au po'o mea matautia.
2.1 Fa'aiti'itia feso'ota'iga ma vaila'au. Fai ni mea e puipui ai le tagata lava ia pe a tagofia vaila'au (fa'ata'ita'iga, matatioata saogalemu, totigilima, po'o lavalava puipui).
2.2 Fa'aiti'itia le inuina o vaila'au. Aua ne'i tatala pusa vaila'au. Fa'aoga na'o le sa'o lelei o le ea (mo se fa'ata'ita'iga, fa'aulu asu).
2.3 Siaki i taimi uma mo vaila'au liki po'o masa'a. Afai e tupu se liki po'o se masaa, mulimuli i faiga fa'amama a le gaosi oloa e pei ona fautuaina i le SDS.
2.4 Taulima otaota vaila'au i totonu o se pusa asu.
2.5 Fa'amautinoa le fa'aogaina o pusa lapisi muamua ma le lua. (O se otaota muamua
koneteina o lo'o taofia ai le otaota vave. O se atigi pusa lona lua o lo'o i ai masaa po'o tafe mai le pusa muamua. O atigipusa uma e lua e tatau ona fetaui ma mea otaota ma fetaui ma feterale, setete, ma le lotoifale mo le teuina o pusa.).
2.6 A uma ona sasaa se atigipusa otaota, faʻamaufaʻailoga i le pulou ua saunia.
2.7 Fa'ailoga (e ala i au'ili'ili pe a mana'omia) otaota e fa'atupuina e fa'aoga fa'apitoa, reagents, ma mea e fa'aogaina i lau falesu'esu'e.
2.8 Fa'amautinoa o lo'o teuina, fa'aliliu, felauaiga, ma lafoa'i otaota e tusa ai ma tulafono fa'alotoifale, setete/provini, ma/po'o le atunu'u.
2.9 Leitio po'o mea fa'aletino e ono mana'omia le tagofiaina fa'apitoa, ma e ono fa'agata ai le lafoa'iina.
3.Biological lamatiaga saogalemu
Avanoa e ono lamatia. Fa'alagolago i fa'ata'ita'iga o lo'o fa'aogaina i luga ole meafaifa'aili, e mafai ona ta'ua le pito i luga ole biohazard. Fa'aaogā metotia fa'amama talafeagai pe a galulue fa'atasi ma meaola fa'aola.
BIOHAZARD. O fa'ata'ita'iga fa'aola e pei o a'a, sua o le tino, vaila'au fa'ama'i, ma le toto o tagata ma isi manu e mafai ona feavea'i fa'ama'i pipisi. Mulimuli i tulafono fa'apitonu'u uma, setete/provini, ma/po'o le atunu'u. Fai mea faigaluega puipui talafeagai, e aofia ai ae le gata i: mata puipui, talipupuni mata, ofu/ofu falesuesue, ma totigilima. O galuega uma e tatau ona faia i totonu o fale fa'apipi'i lelei e fa'aaoga ai meafaigaluega mo le saogalemu (fa'ata'ita'iga, masini fa'apipi'i tino). E tatau ona a'oa'oina tagata ta'ito'atasi e tusa ai ma tulafono fa'atonutonu ma mana'oga a le kamupani/fa'alapotopotoga a'o le'i galulue i mea e ono a'afia ai.
Faitau ma mulimuli i taʻiala talafeagai ma/poʻo manaʻoga faʻatonutonu i mea nei:
I le US: US Department of Health and Human Services guidelines lolomi i Biosafety in Microbiological and Biomedical Laboratories maua i: www.cdc.gov/biosafety.
Tulaga o le Saogalemu ma le Soifua Maloloina, Fa'ama'i Fa'atoto (29 CFR§1910.1030), maua i:
www.access.gpo.gov/nara/cfr/waisidx_01/29cfr1910a_01.html
Polokalama Biosafety a lau kamupani/fa'alapotopotoga mo le galue ma/taulimaina o mea e ono a'afia ai. O lo'o maua fa'amatalaga fa'aopoopo e uiga i ta'iala ole biohazard ile: www.cdc.gov.
I totonu ole EU: Siaki ta'iala fa'alotoifale ma tulafono ile puipuiga ole biosafety ma biosafety ma fa'asino ile faiga sili ona lelei na lomia ile World Health Organization (WHO) Laboratory Biosafety Manual, lomiga lona tolu, maua ile: www.who.int/csr/resources/publications /biosafety/WHO_CDS_CSR_LYO_200 4_11/en/.
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