Buccal Swab/FTA Kadhi DNA Isolation Kit Genomic DNA Kubvisa kana Purifiation Kit kubva kuBuccal Swabs
Kit Description:
Kurumidza kuchenesa yemhando yepamusoro genomic DNA kubva kune buccal swab/FTA Kadhi samples.
◮Hapana RNase kusvibiswa:DNA-Chete Column yakapihwa nekiti inoita kuti zvibvire kubvisa RNA kubva mujenomic DNA pasina kuwedzera RNase panguva yekuyedza, kudzivirira rabhoritari kuti isasvibiswa neRNase yekunze.
◮KumhanyaForegene Protease ine chiitiko chepamusoro kupfuura maprotease akafanana, uye inogaya masampula enyama nekukurumidza;kuvhiya kuri nyore, uye genomic DNA yekubvisa oparesheni inogona kupedzwa mukati me20-80 maminetsi.
◮Yakanakira:Iyo centrifugation inoitwa pakamuri tembiricha, uye hapana chikonzero che4 ° C yakaderera-tembiricha centrifugation kana ethanol kunaya kweDNA.
◮Safety:Hapana organic reagent extract inodiwa.
◮Hunhu hwepamusoro:Iyo yakabviswa genomic DNA ine zvimedu zvakakura, hapana RNA, hapana RNase, uye yakanyanya kuderera ion yemukati, inogona kusangana nezvinodiwa zvekuedza kwakasiyana.
◮Micro-elution system:Inogona kuwedzera kuwanda kweiyo genomic DNA, iyo yakanakira kudzika kwerwizi kana kuyedza.
Tsanangudzo
Iyi kit inopa inoshanda uye inokurumidza nzira yekuwana yakakwirira-concentration genomic DNA kubva ku buccal swabs uye FTA Card (makwapa eropa).Kushandisa kambani yedu's yakasarudzika DNA-chete silica membrane spin column uye formula, yakasanganiswa neForegene Protease, yakakwirira-concentration, yepamusoro-mhando genomic DNA inogona kutorwa mu80 maminetsi.Iyo yakanyatsogadzirirwa diki yekunatsa koramu inosunga genomic DNA, uye iyo DNA inogona kuburitswa nemari shoma (15μl) elution sisitimu yekuwedzera kuwanda kweiyo yakawanikwa genomic DNA, iri nyore pakuona pasi pemvura kana kuyedza.Iyo kit inogona kugadzirisa imwe kana mamwe sampuli panguva, uye nzira yekuchenesa haidi kubviswa kwezvinhu zvakasikwa zvakadai se phenol, chloroform, uye nguva-inopedza isopropanol kana ethanol kunaya, uye kushanda kuri nyore uye kuchengetedza nguva.
Zvinotsanangurwa
50 Preps
Kit zvikamu
| Buffer ST1 |
| Buffer ST2 |
| Linear Acrylamide |
| Buffer PW |
| Buffer WB |
| Buffer EB |
| Foregene Protease |
| DNA-Chete Column |
| Mirayiridzo |
Zvimiro&zvakanakira
-Kwete kusvibiswa kweRNase: Iyo DNA-Chete Column yakapihwa nekiti inoita kuti zvibvire kubvisa RNA kubva kune genomic DNA pasina kuwedzera RNase panguva yekuyedza, kudzivirira rabhoritari kuti isasvibiswa neRNase yekunze.
-Kukurumidza kukurumidza: Foregene Protease ine basa rakakwirira kudarika maprotease akafanana, inochera samples nokukurumidza;nyore kushanda.
-Yakanaka: Iyo centrifugation inoitwa pane tembiricha yekamuri, uye hapana chikonzero che4 ° C yakaderera-kupisa centrifugation kana ethanol precipitation yeDNA.
-Kuchengeteka: Hapana organic reagent kutorwa kunodiwa.
-Hunhu hwepamusoro: Iyo yakabviswa genomic DNA ine zvimedu zvakakura, hapana RNA, hapana RNase, uye yakanyanya kuderera ion yemukati, inogona kusangana nezvinodiwa zvekuedza kwakasiyana.
-Micro-elution system: Inogona kuwedzera kuwanda kweiyo genomic DNA, iri nyore pakuona kudzika kwemvura kana kuedza.
Kit application
Inokodzera kucheneswa kwegenomic DNA kubva kune anotevera samples: buccal swabs, FTA Kadhi (mavara eropa).
Storage uye Sherufu upenyu
-Iyi kititi inogona kuchengetwa kwemwedzi gumi nemaviri pasi pemamiriro akaoma pakushambidzika kwekamuri (15-25 ° C);kana ichida kuchengetwa kwenguva yakareba, inogona kuchengetwa pa2-8°C.
Ongorora: Kana yakachengetwa pane tembiricha yakaderera, mhinduro yacho inonangana nekunaya.Usati washandisa, iva nechokwadi chekuisa mhinduro mukiti pane tembiricha yekamuri kwenguva yakati.Kana zvichidikanwa, preheat mubhati yemvura ye 37 ° C kwemaminitsi gumi kuti iparadze mvura, uye isanganise isati yashandiswa.
-Forgene Protease mhinduro ine chimiro chakasiyana-siyana, iyo inoshanda kana yakachengetwa mukamuri yekushisa kwenguva yakareba (mwedzi mitatu);basa rayo uye kugadzikana zvichava nani kana zvakachengetwa pa4 ° C, saka zvinokurudzirwa kuichengeta pa4 ° C, yeuka kuti usaichengeta pa -20 ° C.
Koramu yekuchenesa yakavharwa
Mune kit iyi, mune genomic DNA yekubvisa oparesheni, mbiru yekuchenesa inoshambadzirwa zvakananga pane sampuli enzymatic lysis musanganiswa pasina centrifugation nhanho, uye mbiru yekuchenesa inogona kuvharika nekuda kwekusakwana enzymization uye yakakwirira viscosity yemuenzaniso.
Zvikonzero zvinobvira zvinotevera zvinotevera:
1. Kusakwana enzymatic digestion yemasampuli ematishu.
Kurudziro: Iyo yekuenzanisira yekugadzirisa nguva yeForegene Protease inogona kuwedzerwa zvakafanira kana iyo supernatant inogona kutorwa mushure me centrifugation pa12,000 rpm (~ 13,400 × g) ye5 min.
2. Kunyanya kushandiswa kwematishu samples kana matishu makuru.
Kurudziro: Zvakanakisa kusapfuudza 1 Buccal swab mumuenzaniso;kana sampuli yakakura kwazvo, wedzera dosi yeBuffer ST1, Foregene Protease, buffer ST2 maererano.
3. Muenzaniso viscosity yakanyanyisa.
Kurudziro: Masamples anogona kucheneswa zvakafanira ne10 mM yeTris-HCl isati yatorwa genomic DNA.
4. Zvimedu zvekadhi reropa zvakayamwa.
Kurudziro: Iyo inopfupika centrifugation nguva yenhanho 6 yeropa nzvimbo (FTA Kadhi) genomic extraction inogona kuwedzerwa zvakafanira.
Goho rakaderera kana pasina DNA
Pane kazhinji zvinhu zvakasiyana-siyana zvinokanganisa goho reGenomic DNA, kusanganisira mabviro emuenzaniso, mamiriro ekuchengetedza emuenzaniso, kugadzirira kwemuenzaniso, kunyengedza, nezvimwe.
Genomic DNA haigone kuwanikwa panguva yekubvisa
Zvikonzero zvinobvira ndezvizvi:
1. Kuchengetedzwa kusina kunaka kwemasampuli kana kuchengetwa kwenguva yakareba kunotungamirira kune genomic DNA degradation.
Kurudziro: Oral swabs inofanirwa kuve ichangobva sampuli, uye hazvikurudzirwe kushandisa yakachengetedzwa swabs ye genomic DNA yekubvisa mashandiro;masampu eropa anofanirwa kuve nechokwadi chekuti mhando yacho inokodzera uye nguva yekuchengetedza haifanirwe kunge yakareba.
2. Kunyanya kushandiswa kwetishu kunogona kukonzera kusatorwa kweiyo genomic DNA inoenderana.
Kurudziro: Tevedzera mirairo yebuccal swab sampling mugwaro rekuvhiya, uye pukuta kakawanda sezvinobvira kuitira kuti maseru akakwana agone kusungirirwa pamuromo swab ye genomic DNA kutorwa;pakutorwa kwesampuli yeropa, nzvimbo yekucheka ropa inogona kuwedzerwa zvakakodzera.
3. Foregene Protease yakachengetedzwa zvisina kunaka, zvichiita kuti iderere mukuita kwayo kana kusashanda.
Kurudziro: Simbisa mamiriro ekuchengetedza eForegene Protease kana kuitsiva neForegene Protease itsva kuitira enzymatic reaction.
4. Kuchengetedza kusina kukodzera kwekiti kana nguva yekuchengetedza yakanyanyisa, zvichiita kuti kukundikana kwezvimwe zvikamu mukiti.
Kurudziro: Tenga nyowani yeBuccal swab DNA yekuzviparadzanisa nevamwe yemaitiro ane hukama.
5. Buffer WB haiwedzeri absolute ethanol.
Kurudziro: Simbisa kuti buffer WB inowedzera huwandu chaihwo hweabsolute ethanol.
6. Iyo yakashata haina kunyatsowedzerwa kune silicone film.
Kurudziro: Wedzera 65 ° C pre-yakadziya eluent inodonha kusvika pakati pesilicon membrane uye siya pakamuri tembiricha kwemaminetsi mashanu kuti uwedzere kushanda kwe elution.
Yakaderera-goho genomic DNA yakaparadzaniswa
Zvikonzero zvinobvira zvinotevera zvinotevera:
1. Kuchengetedzwa kusina kunaka kwemasampuli kana kuchengetwa kwenguva yakareba kunotungamirira kune genomic DNA degradation.
Kurudziro: Oral swabs anonyanya kutorwa samples, uye akachengetedzwa swabs haafanirwe kushandiswa kune genomic DNA kutorwa.
2. Kana chiyero chetishu sampuli chiri chidiki, iyo yakabviswa genomic DNA zvinyorwa zvichave zvishoma.
Kurudziro: Tevedzera iyo oral swab sampling rairo mugwaro rekushandisa, uchipukuta kakawanda sezvinobvira kuitira kuti masero akakwana agone kusungirirwa pamuromo swab ye genomic DNA kutorwa.
3. Foregene Protease yakachengetedzwa zvisina kunaka, zvichiita kuti iderere mukuita kwayo kana kusashanda.
Kurudziro: Simbisa mamiriro ekuchengetedza eForegene Protease kana kuitsiva neForegene Protease itsva kuitira enzymatic reaction.
4. Eluent matambudziko.
Kurudziro: Shandisa Buffer EB ye elution;kana uchishandisa ddH2O kana zvimwewo zvinonzwisisika, simbisa kuti pH yeiyo eluate iri pakati pe7.0-8.5.
5. The eluate haina kunyatsowedzerwa dropwise.
Kurudziro: Wedzera madonhwe emvura pakati pesilicone membrane uye siya pakamuri tembiricha kwemaminetsi mashanu kuti uwedzere kushanda kwe elution.
6. The elution liquid inoungana zvishoma.
Kurudziro: Shandisa eluent ye genomic DNA elution sezvinodiwa mumirayiridzo, ingangoita isingasviki 15 μl.
Kuchena kwakaderera kwegenomic DNA kwakaparadzaniswa
Low genomic DNA kuchena kunogona kutungamirira mukukundikana kana kusagutsikana kwemigumisiro yezviyedzo zvepasi, zvakadai se: ma enzymes haagone kuchekwa akazaruka, PCR haigoni kuwana gene fragment yekufarira, nezvimwewo.
Zvikonzero zvinobvira ndezvizvi:
1. Heteroprotein kusvibiswa, RNA kusvibiswa.
Kuongorora: Koramu yekuchenesa haina kugezwa uchishandisa Buffer PW;iyo Buffer PW yekuchenesa koramu haina kugezwa uchishandisa iyo chaiyo centrifugal kumhanya.
Kurudziro: Iva nechokwadi chekuti hapana kunaya musimba repamusoro usati wawedzera ethanol;iva nechokwadi chekushambidza mbiru yekuchenesa maererano nemirairo, uye danho iri harigoni kusiyiwa.
2. Kusvibiswa ion kusvibiswa.
Ongororo: Buffer WB yekugezesa koramu yakasiiwa kana kuwachwa kamwe chete, zvichikonzera kusvibiswa kweionic kwasara.
Kurudziro: Iva nechokwadi chekugeza Buffer WB kaviri sekurairwa kuti ubvise maion akasara zvakanyanya sezvinobvira.
3. RNA enzyme kusvibiswa.
Analysis: Foreign RNases dzakawedzerwa kune buffer;Buffer PW wash operation yanga isiriyo, zvichikonzera RNase zvisaririra, zvichikanganisa maitirwo ekuedza eRNA ezasi, senge in vitro transcription.
Kurudziro: Foregene series nucleic acid isolation kits inogona kubvisa RNA pasina kuwedzera kuwedzera kweRNase,saka buccal Swab/FTA Kadhi DNA Isolation Kit haifaniri kuwedzera RNase;iva nechokwadi chekutevera mirairo yeBuffer PW yekuchenesa mbiru, uye danho iri harigone kusiiwa.
4. Ethanol yakasara.
Ongororo: Buffer WB haina kuita isina chinhu chubhu centrifugation mushure mekugeza koramu yekuchenesa.
Kurudziro: Ita iyo chaiyo isina chinhu chubhu centrifugation oparesheni zvinoenderana nemirairo.
5. Kumwe kusvibiswa kwetsvina.
Ongororo: Samples dzakachengetwa kana akakosha masampuli haana preteated.
Kurudziro: Nyatsofanogadzirira muenzaniso sezvaunenge warairwa.
