■ Tulaga maualuga mama: O le silica membrane e faʻateʻaina le tele o mea leaga.
■ Lautele faʻaaogaina: Fetaui lelei mo aano o sela, sela, toto ma le tele o isi ituaiga o faʻataʻitaʻiga.
■ Faagaioiga vave: Genomic DNA mafai ona maua i totonu o le 1 itula.
Ituaiga: Faʻavaʻa koluma faʻavae
Faʻataʻitaʻiga ma amata voluma: 100-400 μl toto atoa mai mamalia, 5-20 μl toto atoa mai manulele poʻo amphibian, 106-107 sela faʻaleaganuʻu, 30 mg pepa solo
Galue: 3-40 μg
Taimi o faʻagaioiga: ~ 1 itula
Talosaga i lalo
■ Faʻamolemole faʻaaoga fou faʻataʻitaʻiga e mautinoa ai le faʻamaoni o le faʻamamaina gDNA.
■ Afai o le faʻataʻitaʻiga amata aofaʻi sili atu nai lo le tulaga fautuaina o le pusa, e fautuaina e faʻateleina le aofaʻi o Buffer GA ma Buffer GB, ma vaeluaina le koluma utaina i taimi eseese, pe faʻatautaia le faʻataʻitaʻiga faʻaaoga lua viliina koluma.
■ Mo le teuina umi o le gDNA, fautuaina e teu i buffer TE. Faʻamolemole ia mautinoa o le OD260/ FA280 taua o loʻo i le tulaga o le 1.7-1.9, ma le faʻatauaina o gDNA o 100-300 ng / μl.
DNA Gauai mai Faʻataʻitaʻiga Eseese ma Aofai:
Uma oloa mafai ona faʻapitoa mo ODM / OEM. Mo auiliiliga,faʻamolemole kiliki Customized Service (ODM / OEM)
A-1 Maualalo le totoina o sela poʻo siama i le amataga o le faʻataʻitaʻiga - Faʻamalosia le faʻasologa o sela poʻo siama.
A-2 Lysis le lava o faʻataʻitaʻiga - O faʻataʻitaʻiga e leʻi maeʻa maeʻa faʻafefiloi ma le buffer lysis. E fautuaina e fefiloi maeʻa ile pulse-vortexing mo le 1-2 taimi. —Ua le lava le sela o le sela na mafua mai i le faʻaititia o proteinase K. - Le lava le lysis sela poʻo le faʻaleagaina o le polotini ona o le le lava o le vai tavela mafanafana. E fautuaina e tipi le fasi pepa i ni fasi laiti ma faʻalautele le taimi taele e aveʻese uma toega i le lysate.
A-3 Le lava faʻasalalauga DNA. —E leai se ethanol poʻo se maualalo pasene nai lo le 100% ethanol na faʻaopopoina ae le i faʻamatuʻu le lysate i le vilivalavala.
A-4 O le taua ole pH ole faʻaola eletise e maualalo tele. —Faiaso le pH i le va o le 8.0-8.3.
Ethanol totoe i le eluent.
—E i ai toega fufuluina PW i le eluent. O le ethanol e mafai ona aveʻese i le centrifuging le vili pou mo 3-5 min, ona tuʻu ai lea i le potu vevela poʻo le 50 ℃ incubator mo 1-2 min.
A-1 O le faʻataʻitaʻiga e le fou. — Aveʻese se faʻataʻitaʻiga lelei o le DNA e fai ma faʻatonutonu e faʻamautinoa ai pe ua faʻaleagaina le DNA i totonu o le faʻataʻitaʻiga.
A-2 Leaga muamua togafitiga. —Faʻailoaina e le soʻona suavaia oona nitrogen, toe maua le susu, poʻo tele tele o le faʻataʻitaʻiga.
E tatau ona fesuisuiaʻi mea e fai mo faʻataʻitaʻiga eseese. Mo faʻataʻitaʻiga laʻau, ia mautinoa ia olo lelei i le nitrogen suavaia. Mo manu faʻataʻitaʻiga, homogenate poʻo olo lelei i le suavaia oona. Mo faʻataʻitaʻiga ma sela sela e faigata ona gagau, pei o le G + bacteria ma le fefete, e fautuaina e faʻaoga le lysozyme, lyticase poʻo metotia e motusia ai sela o sela.
4992201/4992202 Laʻau Genomic DNA Kit faʻaaogaina se koluma-faʻavae metotia e manaʻomia chloroform mo le aveʻesea. E sili ona talafeagai mo eseese fualaʻau faʻataʻitaʻi, faʻapea foi ma laʻau mago paʻu. O le Hi-DNAsecure Plant Kit o loʻo faʻavaeina foi i luga ole koluma, ae leai se manaʻoga o le faʻateʻaina o le phenol / chloroform, e saogalemu ai ma e le oona foi. E fetaui lelei mo laʻau ma maualuga polysaccharides ma polyphenol mea. 4992709/4992710 DNAquick Plant System faʻaogaina se vai-faʻavae metotia. Phenol / chloroform aveʻesea e le manaʻomia foi. O le faʻamamaina gaioiga e faigofie ma vave ma leai se tapulaʻa mo le amataga amataga aofaʻiga, o lea e mafai ai e tagata faʻaoga ona fetuʻunaʻi le aofaʻi fetuʻunaʻi e tusa ai ma faʻataʻitaʻiga manaʻoga. Tele lapoa o gDNA fasi e mafai ona maua ma maualuga fua.
E mafai ona faʻatino le vailaʻau toto DNA e faʻaaoga ai meaʻai na saunia i nei mea e lua i le naʻo le suia o le maliega faʻapitoa i faʻatonuga faʻapitoa mo le aveʻesea o le toto o le DNA. O le kopi vaivai o le toto clot DNA aveʻesega maliega maliega faʻapitoa mafai ona tuuina atu pe a talosagaina.
Taofi le faʻataʻitaʻiga fou ile 1 ml PBS, saline masani poʻo le buffer TE. Faʻamaeʻa uma homogenize le faʻataʻitaʻiga e se homogenizer ma ao le oso i lalo o le paipa e centrifuging. Tuʻu ese le supernatant, ma resuspend le oso i le 200 μl buffer GA. Ole faʻamamaina ole DNA e mafai ona faia e tusa ma faʻatonuga.
Mo le faʻamamaina o gDNA i plasma, serum ma tino faʻataʻitaʻiga suavai, TIANamp Micro DNA Pusa ua fautuaina. Mo le faʻamamaina o siama gDNA mai serum / faʻataʻitaʻiga plasma, TIANamp Virus DNA / RNA Kit ua fautuaina. Mo le faʻamamaina o siama gDNA mai serum ma faʻataʻitaʻiga plasma, TIANamp Bacteria DNA Pusa ua fautuaina (lysozyme tatau ona aofia ai mo lelei siama). Mo faʻataʻitaʻiga faua, Hi-Swab DNA Pusa ma TIANamp Bacteria DNA Pusa ua fautuaina.
DNAsecure Plant Pusa poʻo DNAquick Plant System e fautuaina mo le aveʻese mai o le genome. Mo le toʻafilemuina o le genome, TIANamp Yeast DNA Kit e fautuaina (lyticase tatau ona saunia e ia lava).
Talu mai lona faavaeina, o lo tatou fale gaosi ua atiina ae muamua lalolagi vasega oloa ma mulimulitaia le mataupu faavae
o lelei muamua. Tatou oloa ua maua sili lelei taʻuleleia i le alamanuia ma valuabletrusty i fou ma tuai tagata faʻatau ..