Type: Spin ikholamu yakavakirwa
Mharidzo: Plasma, serum, isina maseru emuviri mvura, nezvimwe.
Target: Virus RNA.
Kutanga vhoriyamu: ~ 140 μl
Nguva yekushanda: ~ 1 awa
Pasi pemvura kunyorera: RT-PCR / RT-qPCR, NGS raibhurari yekuvaka, enzyme reaction, nezvimwe.
A-1 Kuderera kwemajeri emasero kana hutachiona mumuenzaniso wekutanga
-Pfumisa kuwanda kwemasero kana mavhairasi.
A-2 Yakaringana lysis yemasampuli
-Iyo mienzaniso haina kusanganiswa zvakakwana ne lysis buffer. Izvo zvinokurudzirwa kunyatso sanganisa ne pulse-vortexing kwe 1-2 nguva.
-Sina kukwana cell lysis inokonzerwa nebasa kuderera kweproteinase K.
-Hakwane cell lysis kana protein degradation nekuda kwekushomeka kwekushambidza nguva 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.
Iyo genomic DNA yakaburitswa kubva kumavhoriyamu akasiyana eropa rese revanhu neTIANamp Ropa reDNA Kit. Mhedzisiro yacho ndeinotevera. Mhedzisiro yacho yakanyorwa sereferenzi chete, iyo chaiyo mhedzisiro mhedzisiro inoenderana nemamiriro esampuli.
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).
Zvese zvigadzirwa zvinogona kuve zvakagadziriswa ODM / OEM. Kuti uwane mamwe mashoko,ndokumbira ubaye Customized Service (ODM / OEM)
AIV-H5 yakasanganiswa kuita gumi-5-10-8gradients ane Mill-Q mvura. 140 μl sampuro yakaburitswa neTIANamp Virus RNA Kit (DP315-R) uye yakakosha kit kubva kuMutengesi A. Inofananidzwa neMutengesi A, TIANGEN ine yepamusoro mashandiro emushoma wevasungwa sampuro.
RT-qPCR yekuona kit: TIANGEN, FastKing Imwe Nhanho RT-qPCR kit (Ongorora)
RT-qPCR chiridzwa: AB 7500 Yekukurumidza Chaiyo-nguva PCR Sisitimu.
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 ..