■ Kuchena kwakanyanya: Silica membrane inobvisa tsvina zhinji.
■ Kushandisa kwakakura: Inokodzera anticoagulant ropa (EDTA, heparin nezvimwewo), jasi rebuffy uye kugwamba kweropa.
Operation Kuvhiya nekukurumidza: Genomic DNA inogona kuwanikwa mukati meawa rimwe chete.
Type: Spin ikholamu yakavakirwa
Muenzaniso uye wekutanga vhoriyamu: 100-1000 μl ropa rese kubva kumammalia, 5-20 μl ropa rese kubva kushiri kana amphibian, 200-1000 μl kuwanda kweropa
Target: Genomic DNA.
Zvibereko: 3-40 μg
Nguva yekushanda: ~ 1 awa
Maodzanyemba ekushandisa: Kudzivirira enzyme kugaya, PCR ongororo, Maodzanyemba blot, chip kusanganiswa
1. Kuti ubvise zvakasara zveRNA, RNase A mhinduro (100 mg / ml) inoda kuzvigadzirira.
2. Ichi chigadzirwa chakanyoreswa sekirasi I in vitrochirwere reagent neBeijing FDA. Ndokumbirawo ubate TIANGEN kana rumwe ruzivo ruchidikanwa.
Zvese zvigadzirwa zvinogona kuve zvakagadziriswa ODM / OEM. Kuti uwane mamwe mashoko,ndokumbira ubaye Customized Service (ODM / OEM)
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Genomic DNA yakabviswa kubva mumasampuli eropa ichishandisa TIANamp Ropa reDNA Kit. M: TIANGEN Maki D15000. Samples: 200 μl yevanhu anticoagulation ropa neEDTA, EDTA / NaF, heparin uye sodium citrate; buffy jasi kubva ku2 ml ropa; 10 μl yehuku yehuku uye 200 μl gonzo reropa. Kune buffy jasi, 2 μl e100 μl eluates akatakurwa; kune vamwe, 4 μl e100 μl eluates akatakurwa pamugwagwa. Mhedzisiro yemhedzisiro: TIANamp Ropa DNA Kit inogona kushandiswa zvakanyanya mukukwenenzverwa kwepamusoro-soro kweDNA kubva kuvanhu anticoagulants nemishonga yakasiyana siyana pamwe neropa kubva kune akasiyana marudzi. |
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Kufananidza kweiyo genomic DNA yakatorwa kubva ku200 μl anticoagulation neEDTA uchishandisa TIANamp Ropa DNA Kit uye zvigadzirwa zvakafanana kubva kune vamwe vanopa. M: TIANGEN Maki D15000. 4 μl ye100 μl eluates akaiswa munzira imwe neimwe. Mhedzisiro yekuedza: Enzanisa nevamwe vatengesi, TIANamp Ropa DNA Kit inogona kuwana goho repamusoro reropa gDNA. |
A-1 Yakadzika kuwanda kwemasero kana hutachiona mune yekutanga sampuro -Pfumisa kuwanda kwemasero kana mavhairasi.
A-2 Kukwana kukosheswa kwemasampuli -Iyo mienzaniso haina kusanganiswa zvakaringana neye lysis buffer. Izvo zvinokurudzirwa kunyatso sanganisa ne pulse-vortexing kwe 1-2 nguva. —Iyo isina kukwana cell lysis inokonzerwa neicho chiitiko kuderera kweproteinase K. - Isina kukwana cell lysis kana protein degradation nekuda kwekushomeka kwekushambidza kwenguva yekugeza. Zvinokurudzirwa kutema zvidimbu zvidimbu zvidiki uye nekuwedzera nguva yekugeza kubvisa ese masara mu lysate.
A-3 Yakakwana DNA adsorption. -Hapana ethanol kana yakaderera-muzana panzvimbo pe100% ethanol yakawedzerwa isati yadzoserwa kune lysate kune spin column.
A-4 Iyo pH kukosha kwe elution buffer yakanyanya kuderera. - Chinja pH kuti ive pakati pe8.0-8.3.
Residual ethanol mune iyo eluent.
—Pane zvakasara zvekuwacha buffer PW mune yakasviba. Iyo ethanol inogona kubviswa ne centrifuging iyo spin ikholamu ye 3-5 min, uye nekuisa pakamuri tembiricha kana 50 ℃ incubator ye 1-2 min.
A-1 Muenzaniso hausi mutsva. -Tora muenzaniso wakanaka weDNA senzira yekuona kuti DNA iri musampu yanyadzisa here.
A-2 Isina kunaka pre-kurapwa. -Yakakonzerwa nekuwandisa kwemvura senitrogen kukuya, hunyoro hunowanikwa, kana huwandu hwakanyanya hwemuenzaniso.
Iwo ekufanofanirwa anofanira kusiyana pamasampuli akasiyana. Pamasampuli emiti, ita shuwa yekukuya zvakakwana muinitrogen yemvura. Kune sampuli yemhuka, homogenate kana kunyatso kukuya muinitrogen yemvura. Kune masampuli ane maseru maseru ayo akaoma kutyora, akadai seG + mabhakitiriya uye mbiriso, zvinokurudzirwa kushandisa lysozyme, lyticase kana mechina nzira kuputsa masero masvingo.
4992201/4992202 Dyara Genomic DNA Kit inotora nzira-yakavakirwa nzira inoda chloroform yekubvisa. Inonyanya kukodzera kune akasiyana masampuli emiti, pamwe neyakaomeswa poda yakaoma. Hi-DNAsecure Plant Kit inoiswawo mumutsetse, asi pasina chikonzero che phenol / chloroform kuburitsa, ichiita kuti ive yakachengeteka uye isiri chepfu. Iyo inokodzera zvirimwa zvine yakakwirira polysaccharides uye polyphenol zvemukati. 4992709/4992710 DNAquick Plant System inotora nzira yemvura-yakavakirwa. Phenol / chloroform kuburitsa hazvidiwe futi. Maitiro ekuchenesa ari nyore uye anokurumidza pasina muganho wemuenzaniso wekutanga, saka vashandisi vanogona kugadzirisa huwandu hwacho nenzira inoenderana nezvinodiwa pakuedza. Kukura kukuru kwezvimedu zveGDNA zvinogona kuwanikwa nezvibereko zvakakura.
Ropa rinovhara ropa reDNA rinogona kuitwa pachishandiswa reagents yakapihwa mumakiti maviri aya nekungochinja protocol kune yakatarwa murau reropa rekuvhara ropa reDNA. Iyo yakapfava kopi yeropa clot DNA yekubvisa protocol inogona kupihwa pakukumbira.
Misa svumbunuro nyowani ne 1 ml PBS, yakasarudzika saline kana TE buffer. Zvakanyatso homogenize muyenzaniso ne homogenizer uye unganidza iyo inodzika pasi pezere pombi ne centrifuging. Dzinga iyo inoshamisa, uye dzorerazve iyo precipitate ne200 μl buffer GA. Iyo inotevera yekunatswa kweDNA inogona kuitwa maererano nemirairo.
Zvekucheneswa kweGDNA muplasma, serum uye muviri fluid sampuro, TIANamp Micro DNA Kit inokurudzirwa. Zvekucheneswa kwehutachiona gDNA kubva kuSerum / plasma sampuro, TIANamp Virus DNA / RNA Kit inokurudzirwa. Zvekucheneswa kwehutachiona gDNA kubva kune serum uye plasma sampuro, TIANamp Bacteria DNA Kit inokurudzirwa (lysozyme inofanirwa kuverengerwa kune yakanaka bhakitiriya). Zvematehwe sampu, Hi-Swab DNA Kit uye TIANamp Bacteria DNA Kit zvinokurudzirwa.
DNAsecure Plant Kit kana DNAquick Plant System inokurudzirwa kubviswa kwefungal genome. Nezve mbiriso genome kuburitsa, TIANamp Yeast DNA Kit inokurudzirwa (lyticase inofanira kuzvigadzirira).
Kubvira payakagadzwa, fekitori yedu yanga ichigadzira zvigadzirwa zvekutanga zvepasirese nekutevedzera mutemo
yemhando yekutanga. Zvigadzirwa zvedu zvakawana mukurumbira wakanaka muindasitiri uye yakakosha kuvimba pakati pevatengi vatsva nevakuru ..