Buccal Swab / FTA Card DNA Isolation Kit Genomic DNA Extraction lossis Purifiaction Kit los ntawm Buccal Swabs
Kit Description:
Ceev ceev purify high-zoo genomic DNA los ntawm buccal swab / FTA daim ntawv kuaj.
◮Tsis muaj RNase contamination:DNA-Tsuas Kem muab los ntawm cov khoom siv ua rau nws tuaj yeem tshem RNA los ntawm genomic DNA yam tsis tau ntxiv RNase thaum lub sij hawm sim, tiv thaiv lub chaw soj nstuam los ntawm exogenous RNase.
◮Ceev ceev:Foregene Protease muaj kev ua haujlwm siab dua li cov protease zoo sib xws, thiab zom cov ntaub so ntswg sai;kev ua haujlwm yog yooj yim, thiab cov genomic DNA rho tawm haujlwm tuaj yeem ua tiav hauv 20-80 feeb.
◮Yooj yim:Lub centrifugation yog ua nyob rau hauv chav tsev kub, thiab tsis tas yuav tsum tau rau 4 ° C low-temperature centrifugation los yog ethanol nag lossis daus ntawm DNA.
◮Kev nyab xeeb:Tsis muaj cov organic reagent extraction yog yuav tsum tau.
◮High quality:Cov genomic DNA uas tau muab rho tawm muaj cov tawg loj, tsis muaj RNA, tsis muaj RNase, thiab cov ntsiab lus tsis tshua muaj ion, uas tuaj yeem ua tau raws li qhov yuav tsum tau muaj ntawm ntau yam kev sim.
◮Micro-elution system:Nws tuaj yeem ua kom cov concentration ntawm genomic DNA, uas yooj yim rau kev tshawb nrhiav los yog kev sim.
Kev piav qhia
Cov khoom siv no muab txoj hauv kev ua tau zoo thiab nrawm kom tau txais cov txiaj ntsig zoo ntawm DNA ntawm buccal swabs thiab FTA Card (cov ntshav me me).Siv peb lub tuam txhab's cim DNA-tsuas yog silica membrane spin kem thiab mis, ua ke nrog Foregene Protease, siab-concentration, zoo genomic DNA tuaj yeem muab rho tawm hauv 80 feeb.Qhov tshwj xeeb tsim me me purification kem khi genomic DNA, thiab DNA tuaj yeem eluted nrog me me (15μl) elution system kom nce cov concentration ntawm cov genomic DNA, uas yog yooj yim rau downstream nrhiav los yog sim.Cov khoom siv tuaj yeem ua tiav ib lossis ntau qhov qauv ntawm ib lub sijhawm, thiab cov txheej txheem purification tsis tas yuav rho tawm cov organic tshuaj xws li phenol, chloroform, thiab siv sij hawm siv isopropanol lossis ethanol nag lossis daus, thiab kev ua haujlwm yooj yim thiab txuag sijhawm.
Specifications
50 Preps
Cov khoom siv
| Buffer ST1 |
| Tsis Siv Neeg ST2 |
| Linear Acrylamide |
| Buffer PW |
| Buffer WB |
| Buffer IB |
| Foregene Protease |
| DNA-Tsuas Kem |
| Cov lus qhia |
Nta & qhov zoo
-Tsis muaj RNase kab mob: Cov DNA-Tsuas Kab uas muab los ntawm cov khoom siv ua rau nws muaj peev xwm tshem RNA los ntawm genomic DNA yam tsis tau ntxiv RNase thaum lub sij hawm sim, zam lub chaw kuaj los ntawm exogenous RNase.
- Ceev ceev: Foregene Protease muaj kev ua haujlwm siab dua li cov proteases zoo sib xws, zom cov qauv sai;kev ua haujlwm yooj yim.
- Yooj yim: Lub centrifugation yog ua nyob rau hauv chav tsev kub, thiab tsis tas yuav tsum tau rau 4 ° C qis-kub centrifugation los yog ethanol nag lossis daus ntawm DNA.
-Kev nyab xeeb: Tsis tas yuav muaj cov organic reagent extraction.
-High zoo: Cov genomic DNA rho tawm muaj cov tawg loj, tsis muaj RNA, tsis muaj RNase, thiab cov ntsiab lus tsis tshua muaj ion, uas tuaj yeem ua tau raws li qhov yuav tsum tau muaj ntawm ntau yam kev sim.
-Micro-elution system: Nws tuaj yeem ua kom cov concentration ntawm genomic DNA, uas yooj yim rau kev tshawb nrhiav los yog kev sim.
Daim ntawv thov Kit
Nws yog qhov tsim nyog rau kev ua kom huv ntawm genomic DNA los ntawm cov qauv hauv qab no: buccal swabs, FTA Card (ntshav stains).
Cia thiab Txee lub neej
-Cov khoom siv no tuaj yeem khaws cia rau 12 lub hlis nyob rau hauv qhuav qhuav ntawm chav tsev kub (15-25 ° C);Yog tias nws xav tau khaws cia rau lub sijhawm ntev, nws tuaj yeem khaws cia ntawm 2-8 ° C.
Lus Cim: Yog tias khaws cia ntawm qhov kub thiab txias, qhov kev daws teeb meem yuav ua rau nag lossis daus.Ua ntej siv, nco ntsoov muab cov tshuaj tso rau hauv cov khoom siv ntawm chav tsev kub rau ib lub sijhawm.Yog tias tsim nyog, preheat nws hauv 37 ° C dej da dej rau 10 feeb kom yaj cov dej nag, thiab sib tov ua ntej siv.
-Foregene Protease tov muaj ib tug tshwj xeeb mis, uas yog active thaum khaws cia nyob rau hauv chav tsev kub ntev (3 lub hlis);nws cov dej num thiab kev ruaj ntseg yuav zoo dua thaum khaws cia ntawm 4 ° C, yog li nws pom zoo kom khaws cia ntawm 4 ° C, nco ntsoov tsis txhob khaws cia ntawm -20 ° C.
Lub purification kem yog clogged
Nyob rau hauv cov khoom no, nyob rau hauv lub genomic DNA extraction ua hauj lwm, lub purification kem yog ncaj qha adsorbed ntawm cov qauv enzymatic lysis sib tov yam tsis muaj centrifugation kauj ruam, thiab purification kem yuav raug thaiv vim tsis tiav enzymization thiab siab viscosity ntawm cov qauv.
Cov ua tau hauv qab no yog raws li nram no:
1. Tsis tiav enzymatic digestion ntawm cov ntaub so ntswg kuaj.
Kev pom zoo: Lub sijhawm ua piv txwv ntawm Foregene Protease tuaj yeem txuas ntxiv kom tsim nyog lossis tus supernatant tuaj yeem siv tom qab centrifugation ntawm 12,000 rpm (~ 13,400 × g) rau 5 min.
2. Kev siv cov ntaub so ntswg ntau dhau los yog cov ntaub so ntswg loj.
Kev pom zoo: Nws yog qhov zoo tshaj kom tsis pub tshaj 1 Buccal swab hauv cov qauv;Yog tias tus qauv loj dhau, nce qhov ntau npaum ntawm Buffer ST1, Foregene Protease, tsis ST2 raws li.
3. Cov qauv viscosity siab dhau lawm.
Kev pom zoo: Cov qauv tuaj yeem tsim nyog diluted nrog 10 mM ntawm Tris-HCl ua ntej genomic DNA rho tawm.
4. Cov seem ntawm daim npav ntshav tau nqus.
Kev pom zoo: Lub sijhawm centrifugation ib ntus ntawm kauj ruam 6 ntawm ntshav chaw (FTA Card) genomic extraction tuaj yeem txuas ntxiv kom tsim nyog.
Tsawg tawm los lossis tsis muaj DNA
Muaj ntau ntau yam uas cuam tshuam rau genomic DNA yield, nrog rau cov qauv hauv paus chiv keeb, cov qauv khaws cia, qauv npaj, kev tswj xyuas, thiab lwm yam.
Genomic DNA tsis tuaj yeem tau txais thaum lub sijhawm rho tawm
Cov ua tau yog raws li nram no:
1. Kev khaws cia tsis raug ntawm cov qauv lossis khaws cia ntev dhau los ua rau genomic DNA degradation.
Cov lus pom zoo: Lub qhov ncauj swabs yuav tsum zoo dua yog cov qauv tshiab, thiab nws tsis pom zoo kom siv cov swabs khaws cia rau kev ua haujlwm ntawm DNA genomic extraction;Cov ntshav kuaj ntshav yuav tsum xyuas kom meej tias qhov zoo yog tsim nyog thiab lub sijhawm cia yuav tsum tsis txhob ntev dhau.
2. Kev siv cov ntaub so ntswg tsawg dhau yuav ua rau tsis muaj kev rho tawm ntawm cov DNA sib txuas.
Cov lus pom zoo: Ua raws li cov lus qhia ntawm buccal swab sampling hauv cov lus qhia ua haujlwm, thiab so kom ntau li ntau tau kom cov hlwb txaus tuaj yeem txuas rau lub qhov ncauj swab rau genomic DNA extraction;rau cov ntshav qhov chaw coj mus kuaj, qhov chaw txiav cov ntshav tuaj yeem tsim nyog ntxiv.
3. Foregene Protease yog khaws cia tsis raug, ua rau txo qis hauv nws txoj haujlwm lossis kev ua haujlwm tsis zoo.
Kev Pom Zoo: Txheeb xyuas qhov chaw cia ntawm Foregene Protease lossis hloov nrog Foregene Protease tshiab rau cov tshuaj tiv thaiv enzymatic.
4. Kev khaws cia tsis raug ntawm cov khoom siv lossis lub sijhawm khaws cia ntev dhau lawm, ua rau qee yam khoom siv hauv cov khoom siv tsis ua haujlwm.
Kev pom zoo: Yuav ib lub Buccal swab DNA cais cov khoom tshiab rau cov txheej txheem cuam tshuam.
5. Buffer WB tsis ntxiv ethanol kiag li.
Kev pom zoo: Paub meej tias tsis WB ntxiv qhov tseeb ntim ntawm ethanol.
6. Lub eluent tsis raug ntxiv rau cov yeeb yaj kiab silicone.
Kev pom zoo: Ntxiv 65 ° C ua ntej sov eluent tee rau nruab nrab ntawm silicone membrane thiab tawm hauv chav sov li 5 min kom nce qhov kev ua tau zoo.
Tsawg-yield genomic DNA cais
Cov ua tau hauv qab no yog raws li nram no:
1. Kev khaws cia tsis raug ntawm cov qauv lossis khaws cia ntev dhau los ua rau genomic DNA degradation.
Daim ntawv pom zoo: Cov qhov ncauj qhov ncauj yog qhov zoo dua tshiab coj mus kuaj, thiab khaws cia swabs yuav tsum tsis txhob siv rau kev rho tawm DNA genomic.
2. Yog tias cov qauv ntawm cov ntaub so ntswg me me dhau lawm, cov ntsiab lus ntawm DNA genomic extracted yuav tsawg dua.
Kev pom zoo: Ua raws li cov lus qhia ntawm qhov ncauj swab sampling nyob rau hauv phau ntawv qhia kev khiav hauj lwm, so kom ntau li ntau zaus kom cov hlwb txaus tuaj yeem txuas rau lub qhov ncauj swab rau genomic DNA extraction.
3. Foregene Protease yog khaws cia tsis raug, ua rau txo qis hauv nws txoj haujlwm lossis kev ua haujlwm tsis zoo.
Kev Pom Zoo: Txheeb xyuas qhov chaw cia ntawm Foregene Protease lossis hloov nrog Foregene Protease tshiab rau cov tshuaj tiv thaiv enzymatic.
4. Muaj teeb meem.
Kev pom zoo: Siv Buffer EB rau elution;yog siv ddH2O lossis lwm yam eluents, paub meej tias pH ntawm eluate nyob nruab nrab ntawm 7.0-8.5.
5. Lub eluate tsis raug ntxiv dropwise.
Kev pom zoo: Ntxiv eluent tee rau hauv nruab nrab ntawm silicone membrane thiab tawm hauv chav sov li 5 min kom nce elution efficiency.
6. Cov kua elution accumulates tsawg dhau.
Kev pom zoo: Siv eluent rau genomic DNA elution raws li xav tau nyob rau hauv cov lus qhia, yam tsawg kawg nkaus tsis pub tsawg tshaj 15 μl.
Tsawg purity ntawm genomic DNA cais
Tsawg genomic DNA purity tuaj yeem ua rau tsis ua haujlwm lossis tsis txaus siab ntawm cov kev sim hauv qab, xws li: enzymes tsis tuaj yeem txiav qhib, PCR tsis tuaj yeem tau txais cov noob ntawm kev txaus siab, thiab lwm yam.
Cov ua tau yog raws li nram no:
1. Heteroprotein pollution, RNA kuab paug.
Kev tsom xam: Cov kab purification tsis tau ntxuav siv Buffer PW;Buffer PW ntxuav purification kem tsis tau ntxuav siv qhov tseeb centrifugal ceev.
Kev pom zoo: xyuas kom meej tias tsis muaj nag lossis daus hauv lub supernatant ua ntej ntxiv ethanol;nco ntsoov ntxuav lub purification kem raws li cov lus qhia, thiab cov kauj ruam no tsis tuaj yeem raug tshem tawm.
2. impurity ion muaj kuab paug.
Kev tshuaj ntsuam xyuas: Buffer WB ntxuav purification kem raug tshem tawm lossis ntxuav tsuas yog ib zaug, ua rau cov khoom seem ionic.
Kev pom zoo: Nco ntsoov ntxuav Buffer WB 2 zaug raws li qhia kom tshem tawm cov ions seem ntau li ntau tau.
3. RNA enzyme paug.
Kev tsom xam: Cov RNases txawv teb chaws tau ntxiv rau qhov tsis muaj;Buffer PW ntxuav kev ua haujlwm tsis raug, ua rau RNase residues, cuam tshuam rau hauv qab RNA kev sim ua haujlwm, xws li hauv vitro transcription.
Kev pom zoo: Foregene series nucleic acid cais cov khoom siv tuaj yeem tshem tawm RNA yam tsis muaj qhov sib ntxiv ntawm RNase, yog li buccal Swab / FTA Card DNA Isolation Kit tsis tas yuav ntxiv RNase;Nco ntsoov ua raws li cov lus qhia rau Buffer PW ntxuav purification kem, thiab cov kauj ruam no tsis tuaj yeem raug tshem tawm.
4. Ethanol residue.
Kev tshuaj xyuas: Buffer WB tsis ua qhov khoob raj centrifugation tom qab ntxuav lub purification kem.
Kev pom zoo: Ua qhov raug khoob raj centrifugation ua haujlwm raws li cov lus qhia.
5. Lwm yam impurity muaj kuab paug.
Kev Ntsuam Xyuas: Cov qauv khaws tseg lossis cov qauv tshwj xeeb tsis raug ua ntej.
Kev pom zoo: Ua tib zoo pretreat tus qauv raws li qhia.
