Lub tswv yim rau kev txhim kho cov kua nplaum rov qab

1. Ua kom cov qauv load thaum lub sij hawm electrophoresis.

2. Siv cov khoom tshiab tshiab uas tsis muaj electrophoresis.

3. Thaum txiav cov kua nplaum, sim txiav cov kua nplaum nkaus xwb nrog cov strips kom txo cov ntim ntawm cov kua nplaum txiav: tsis tas yuav cov kua nplaum nrog ob peb lub hom phiaj tawg, txwv tsis pub nws yuav cuam tshuam tus nqi rov qab.

4. Tom qab melting ob los yog ntau daim ntawm cov kua nplaum, siv ib lub raj txawm tias lub ntim loj npaum li cas thiab hloov mus rau tib kem.

5. Cov tshuaj ntxiv rau hauv cov sol tuaj yeem ua rau me ntsis ntxiv, uas yog qhov zoo rau kev khi ntawm DNA rau daim nyias nyias, tab sis feem ntau tsis tshaj 750ul.

6. Tus yuam sij rau gel rov qab yog khi DNA rau hauv kem los ntawm cov ntsev concentration, acidity (them) thiab hydrophobicity ntawm cov tshuaj nyob rau hauv kem.Yog li ntawd, yog tias pH ntawm lub electrophoresis tsis yog siab dhau, 10ul (pH 5.0, 3mol / L NaAC) tuaj yeem muab ntxiv rau lub sol;txhawm rau txhawm rau cuam tshuam DNA molecules zoo dua ntawm daim nyias nyias, 30% isopropanol tuaj yeem muab ntxiv rau hauv cov kua tom qab yaj cov kua nplaum.

7. Ua ntej ntxiv cov eluent, tawm hauv kem ntawm chav tsev kub li ob peb feeb (kwv yees li 10 feeb) kom tag nrho cov ethanol evaporate.

8. Thaum kawg, ntxiv tsawg eluent kom txo qhov ntim rov qab.Feem ntau, 30-50μl eluent yog siv rau elution (tsis yog tsawg heev, txwv tsis pub nws yuav tsis tuaj yeem ntub cov membrane, uas tsis haum rau elution);cov elution droplets yog nyob rau hauv nruab nrab ntawm daim nyias nyias, kom tag nrho elute DNA khi rau daim nyias nyias.

9. Tom qab ntxiv cov eluent, nws tuaj yeem eluted hauv dej da dej ntawm 55 degrees rau 5 feeb los yog muab tso rau hauv dej da dej ntawm 50 degrees rau ntau tshaj 10 feeb, los yog kaw nrog parafilm ntawm 4 degrees thaum hmo ntuj, thiab tom qab ntawd centrifuged rov qab rau hnub tom qab, cov nyhuv zoo.

10.Ntxiv cov centrifuged eluate rov qab mus rau sab adsorption thiab centrifuge dua.

Cov txheej txheem ntxaws thiab cov txheej txheem rau PCR cov khoom rov qab

1. Kev siv roj hmab zoo tib yam

Yog tias koj xav rov qab cov kua nplaum, nws yog qhov zoo tshaj plaws los siv cov khoom siv, uas yog qhov yooj yim thiab muaj qhov rov zoo dua me ntsis.Yog tias koj xav tau rov qab los ntawm nws tus kheej, koj tuaj yeem ntxiv 3 npaug ntawm TE tom qab txiav cov kua nplaum.Tom qab yaj hauv dej da dej, phenol, phenol / chloroform raug muab rho tawm kom huv, thiab ethanol yog precipitated.Qhov ntawd yog nws.

2. DNA rov qab los ntawm qis melting point gels

Purification ntawm DNA Fragments Ntxiv TE (10 mmol / l Tris-HCl pH8.0, 0.1 mmol / l EDTA) sib npaug rau cov ntim ntawm cov gel, thiab tso rau hauv 65 ° C dej da dej rau 5 feeb kom yaj cov gel kom tiav.

Tom qab nws raug coj mus rau hauv chav tsev kub, qhov sib npaug ntawm phenol (saturated nrog TE, TE tau muab ntim rau hauv txheej sab saud, thiab cov txheej txheem qis ntawm phenol raug tshem tawm) tau ntxiv, thiab cov sib tov tau maj mam sib xyaw (tsis tas yuav tsum tau sib tov), ​​thiab centrifuged ntawm 12,000 rpm rau 3 feeb.Rov ua 1-2 zaug.

Noj cov tshuaj supernatant, ntxiv 0.1 ntim ntawm 3mol / L sodium acetate (pH 5.2) thiab 2.5 npaug ntawm qhov ntim ntawm ethanol kom nqa tawm ethanol nag lossis daus.Dissolve lub purified DNA nrog ib tug tsim nyog tus nqi ntawm TE, ntsuas cov ntsiab lus, thiab npaj rau kev siv (nws yuav siv tau rau lub hom phiaj gene qauv tsom xam, soj ntsuam kev npaj, thiab lwm yam).

3. PCR rov qab nrog qhov zoo amplification tshwj xeeb

Yog tias qhov tshwj xeeb ntawm PCR amplification yog qhov zoo, nws tsuas yog ib qho yooj yim purification thiab rov qab los ntawm PCR cov khoom.Koj tuaj yeem ntxiv 50ug / ml proteinase K rau cov khoom PCR, 37 degrees rau 1h, rho tawm ib zaug nrog phenol / chloroform, rho tawm ib zaug nrog chloroform, thiab ntxiv 0.1 ntim ntawm supernatant.Cov sodium acetate tau zoo los ntawm nag lossis daus nrog 2.5 ntim ntawm ethanol kiag li.

Yam khoom:

https://www.foreivd.com/pcr-purification-kit-2-product/

https://www.foreivd.com/blood-superdirecttm-pcr-kit-edta-product/