I-TGuide Plasma DNA yokuSusa iKhithi (1.2ml)

A-1 Ukuxinana kweeseli okanye intsholongwane kwisampulu yokuqala -Tyebisa uxinzelelo lweeseli okanye iintsholongwane.

A-2 I-lysis engonelanga yeesampulu -Iisampulu azange zixutywe kakuhle kunye ne-lysis buffer. Kucetyiswa ukuba udibanise ngokucofa i-vortexing kangangezihlandlo ezi-1-2. -I-lysis yeseli engonelanga ebangelwe kukuncipha kwemisebenzi ye-proteinase K. Kucetyiswa ukuba usike izicwili zibe ziingceba kwaye wandise ixesha lokuhlamba ukususa yonke intsalela yelysate.

A-3 Ukungoneliseki kwe-DNA adsorption. -Ayikho i-ethanol okanye ipesenti esezantsi endaweni ye-100% ye-ethanol eyongezwe ngaphambi kokuba kudluliselwe i-lysate kwikholamu yokujikeleza.

A-4 Ixabiso le-pH ye-elution buffer liphantsi kakhulu. Lungisa i-pH iye phakathi kwe-8.0-8.3.

Intsalela ye-ethanol kwi-eluent.

-Kukho intsalela yokuhlambela i-PW kwisikali. I-ethanol ingasuswa ngokufaka phakathi kwikholamu ejikelezayo ye-3-5 min, kwaye emva koko ubeke kubushushu begumbi okanye kwi-50 ℃ incubator ye-1-2 min.

A-1 Isampulu ayisiyonto intsha. -Khupha isampulu yeDNA elungileyo njengokulawula ukumisela ukuba ingaba iDNA ekwisampulu ihlaziyiwe.

Unyango lwangaphambi kokunyanga okungalunganga. -Yenziwe kukugaya i-nitrogen engamanzi ngokugqithisileyo, ukubuyisa ukufuma, okanye isixa esikhulu sesampulu.

Ukufakwa kwangaphambili kufuneka kwahluka kwiisampulu ezahlukeneyo. Kwiisampulu zezityalo, qiniseka ukuba usile kakuhle kwi-nitrogen engamanzi. Kwiisampulu zezilwanyana, homogenate okanye ugaye kakuhle kwi-nitrogen engamanzi. Kwiisampulu ezineendonga zeseli ekunzima ukuziqhekeza, ezinjenge-G + bacteria kunye nemvubelo, kucetyiswa ukuba kusetyenziswe i-lysozyme, i-lyticase okanye iindlela zoomatshini zokwaphula iindonga zeseli.

I-4992201/4992202 ye-Genomic DNA Kit isebenzisa indlela esekwe kwikholamu efuna ikloroform yokukhupha. Ifanelekile ngakumbi kwiisampulu ezahlukeneyo zezityalo, kunye nomgubo owomileyo wezityalo. I-Hi-DNAsecure Plant Kit nayo isekwe kwikholamu, kodwa kungekho mfuneko yokukhutshwa kwe-phenol / chloroform, ukuyenza ikhuseleke kwaye ingabinetyhefu. Kufanelekile ukuba izityalo ezine-polysaccharides ephezulu kunye nomxholo we-polyphenol. 4992709/4992710 DNAquick Plant System yamkela indlela esekwe kulwelo. Ukukhutshwa kwePhenol / chloroform akufuneki nako. Inkqubo yokuhlanjululwa ilula kwaye iyakhawuleza ngaphandle komda kwiisampulu zezixa zokuqala, ke abasebenzisi banokuhlengahlengisa isixa ngokuhambelana neemfuno zovavanyo. Ubungakanani obukhulu bamaqhekeza e-gDNA anokufunyanwa ngesivuno esikhulu.

I-genomic DNA yakhutshwa kwimilinganiselo eyahlukeneyo yeesampulu zabantu zegazi yi-TIANamp Blood DNA Kit. Iziphumo zilandelayo. Iziphumo zidweliswe njengereferensi kuphela, ezona ziphumo zokukhupha zixhomekeke kwimeko yeesampulu.

Ukukhutshelwa kwegazi le-DNA kungena kusetyenziswa ii-reagents ezibonelelwe kwezi khithi zimbini ngokutshintsha nje umthetho olandelwayo kumyalelo othile wokukhutshwa kwegazi le-DNA. Ikopi ethambileyo yenkqubo yokutsalwa kwegazi kwi-DNA inokukhutshwa xa uceliwe.

Misa isampulu entsha nge-1 ml ye-PBS, isaline eqhelekileyo okanye i-TE buffer. Ngokupheleleyo homogenize isampuli yi homogenizer kwaye uqokelele isitshisi ukuya ezantsi kumbhobho nge-centrifuging. Yilahle into engaphaya kwamandla, kwaye uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde uphinde wenziwe khona nge-200 μl buffer GA. Ukucocwa kwe-DNA elandelayo kunokwenziwa ngokomyalelo.

Ukucoca i-gDNA kwi-plasma, i-serum kunye neesampuli zomzimba, kuyacetyiswa i-TIANamp Micro DNA Kit. Ukucoca intsholongwane gDNA kwiisampulu ze-serum / plasma, I-TIANamp Virus DNA / RNA Kit iyacetyiswa. Ukucoca i-gDNA yebhaktiriya kwiisampulu ze-serum kunye neplasma, i-TIANamp Bacteria DNA Kit iyacetyiswa (i-lysozyme kufuneka ibandakanywe kubhaktiriya). Kwiisampulu zamathe, iHi-Swab DNA Kit kunye ne-TIANamp Bacteria DNA Kit ziyacetyiswa.

I-DNAsecure Plant Kit okanye i-DNAquick Plant System iyacetyiswa ukuba isuswe kwi-fungal genome extraction. Ukukhutshwa kwegwele le-genome, i-TIANamp Yeast DNA Kit iyacetyiswa (i-lyticase kufuneka izilungiselele).