Zotsatirazi kusanthula mavuto zotheka muBuccalswab/Mtengo FTA card DNA extraction is helpful to your experiment. In addition, for other experimental or technical problems in addition to operating instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us at: 028-83360257 or E-mali:Tech@foregene.com.

Mzere woyeretsa watsekedwa

Mu zida izi, mu genomic DNA m'zigawo ntchito, ndime kuyeretsedwa mwachindunji adsorbed pa chitsanzo enzymatic lysis osakaniza popanda sitepe centrifugation, ndi ndime kuyeretsedwa akhoza oletsedwa chifukwa chosakwanira enzymization ndi mkulu mamasukidwe akayendedwe a chitsanzo.

Zifukwa zotsatirazi ndi izi:

1. Kusakwanira kwa enzymatic chimbudzi cha zitsanzo za minofu.

Malangizo: Zitsanzo zakukonza nthawi ya Foregene Protease zitha kukulitsidwa moyenerera kapena supernatant ingatengedwe pambuyo pa centrifugation pa 12,000 rpm (~ 13,400× g) kwa mphindi 5.

2. Kugwiritsa ntchito kwambiri zitsanzo za minofu kapena minofu yaikulu.

Malangizo: Ndibwino kuti musapitirire 1Buccal swab mu chitsanzo;ngati chitsanzocho ndi chachikulu kwambiri, onjezani mlingo wa Buffer ST1, Foregene Protease, buffer ST2 moyenerera.

3. Kukhuthala kwachitsanzo ndikokwera kwambiri.

Malangizo: Zitsanzo zitha kuchepetsedwa moyenera ndi 10 mM ya Tris-HCl musanayambe kutulutsa ma genomic DNA.

4. Zidutswa za khadi la magazi zayamwa.

Malangizo: Nthawi yocheperako ya centrifugation ya sitepe 6 ya malo amagazi (FTA Card) yotulutsa ma genomic imatha kukulitsidwa moyenerera.

Zokolola zochepa kapena palibe DNA

Nthawi zambiri pamakhala zinthu zosiyanasiyana zomwe zimakhudza zokolola za DNA ya genomic, kuphatikizapo chiyambi cha zitsanzo, mikhalidwe yosungiramo zitsanzo, kukonzekera zitsanzo, kusintha, ndi zina zotero.

Genomic DNA sichingapezeke panthawi yochotsa

Zomwe zingatheke ndi izi:

1. Kusungidwa molakwika kwa zitsanzo kapena kusungidwa kwa nthawi yayitali kumabweretsa kuwonongeka kwa ma genomic DNA.

Malangizo: Masamba a pakamwa amayenera kutsatiridwa mwatsopano, ndipo sikoyenera kugwiritsa ntchito swabs zosungidwa pochotsa ma genomic DNA;zitsanzo za malo a magazi ziyenera kuonetsetsa kuti khalidweli ndi loyenerera ndipo nthawi yosungira sikuyenera kukhala yaitali kwambiri.

2. Kugwiritsa ntchito minofu pang'ono kungapangitse kuti DNA ya ma genomic isatuluke.

Malangizo: Tsatiranibuccal malangizo a zitsanzo za swab mu bukhu la opareshoni, ndi kupukuta kangapo momwe mungathere kuti maselo okwanira athe kumangirizidwa ku swab yapakamwa kuti muchotse DNA ya genomic;pochotsa magazi, malo odulira magazi amatha kuonjezedwa moyenerera.

3. Foregene Protease imasungidwa molakwika, zomwe zimapangitsa kuchepa kwa ntchito yake kapena kusakhazikika.

Malangizo: Tsimikizirani zosungirako zandi Foregene Protease kapena m'malo mwake ndi Foregene Protease yatsopano ya enzymatic reaction.

4. Kusungidwa kosayenera kwa zida kapena nthawi yosungirako ndi nthawi yayitali kwambiri, zomwe zimapangitsa kuti zigawo zina ziwonongeke muzitsulo.

Malangizo: Gulani yatsopanoBuccal DNA yambakudzipatula zida za ndondomeko zogwirizana.

5. Buffer WB sichiwonjezera ethanol.

Malangizo: Tsimikizirani kuti buffer WB imawonjezera voliyumu yolondola ya ethanol.

6. Chosowacho sichinawonjezedwe molondola ku filimu ya silikoni.

Malangizo: Onjezani 65°CMadontho otenthetsera otenthetserapo mpaka pakati pa nembanemba ya silikoni ndikusiya kutentha kwa mphindi 5 kuti muwonjezere mphamvu ya elution.

Low-yield genomic DNA akutali

Zifukwa zotsatirazi ndi izi:

1. Kusungidwa molakwika kwa zitsanzo kapena kusungidwa kwa nthawi yayitali kumabweretsa kuwonongeka kwa ma genomic DNA.

Malangizo: Masamba amkamwa amasankhidwa mwatsopano, ndipo ma swabs osungidwa sayenera kugwiritsidwa ntchito pochotsa ma genomic DNA.

2. Ngati kuchuluka kwa minofu kuli kochepa kwambiri, zomwe zatulutsidwa mu DNA zimakhala zochepa.

Malangizo: Tsatirani malangizo a oral swab mu bukhu la opareshoni, pukutani kangapo momwe mungathere kuti ma cell okwanira amangiridwe pakamwa kuti muchotse DNA ya genomic.

3. Foregene Protease imasungidwa molakwika, zomwe zimapangitsa kuchepa kwa ntchito yake kapena kusakhazikika.

Malangizo: Tsimikizirani zosungirako zathe Foregene Protease kapena m'malo mwake ndi chatsopano Foregene Protease kwa enzymatic reaction.

4. Mavuto osaneneka.

Malangizo: Gwiritsani ntchito Buffer EB kwa elution;ngati mukugwiritsa ntchito ddH₂O kapena zofotokozera zina, zimatsimikizira kuti pH ya eluate ili pakati pa 7.0-8.5.

5. The eluate sanawonjezedwe molondola dropwise.

Malangizo: Onjezani madontho osawoneka bwino pakati pa nembanemba ya silikoni ndikusiya kutentha kwa mphindi 5 kuti muwonjezere mphamvu.

6. Madzi a elution amawunjikana pang'ono.

Malangizo: Gwiritsani ntchito eluent pakuwunika kwa ma genomic DNA monga momwe akufunira mu malangizo, osacheperaosachepera pa 15μl.

Luwu chiyero of genomic DNAakutali

Low genomic DNA chiyero kungayambitse kulephera kapena zotsatira zosakhutiritsa za zoyesera zotsika, monga: ma enzyme sangatsegulidwe, PCR satha kupeza gawo lachidwi, ndi zina zambiri.

Zomwe zingatheke ndi izi:

1. Heteroprotein kuipitsa, RNA kuipitsa.

Kusanthula: Mzere woyeretsedwa sunatsukidwe pogwiritsa ntchito Buffer PW;ndime yoyeretsa ya Buffer PW sinatsukidwe pogwiritsa ntchito liwiro lolondola la centrifugal.

Malangizo: Onetsetsani kuti palibe mvula mumphamvu yamphamvu musanawonjezere Mowa;onetsetsani kuti mukutsuka ndime yoyeretsera molingana ndi malangizo, ndipo sitepe iyi siyingasiyidwe.

2. Kudetsedwa kwa ayoni.

Kuwunika: Mzere woyeretsera wa Buffer WB sunasiyidwe kapena kutsukidwa kamodzi kokha, zomwe zidapangitsa kuipitsidwa kwa ayoni.

Malangizo: Onetsetsani kuti mukutsuka Buffer WB ka 2 monga mwalangizidwa kuti muchotse ma ion otsalira momwe mungathere.

3. Kuwonongeka kwa enzyme ya RNA.

Kusanthula: Ma RNase akunja adawonjezedwa ku buffer;Opaleshoni yotsuka ya Buffer PW inali yolakwika, zomwe zidapangitsa kuti zotsalira za RNase, zomwe zikukhudza zoyeserera zakutsika za RNA, monga zolembera mu vitro.

Malangizo: Foregene mndandanda nucleic acidisolazida zothandizira zimatha kuchotsa RNA popanda kuwonjezera kwa RNase,motero buccal Swab/FTA Card DNA Isolation Kitzofunika't kuwonjezera RNase;onetsetsani kuti mwatsatira malangizo a gawo la kuyeretsedwa kwa Buffer PW, ndipo sitepe iyi siyingasiyidwe.

4. Zotsalira za Ethanol.

Kusanthula: Buffer WB sanachite kanthu chubu centrifugation atatsuka ndime yoyeretsa.

Malangizo: Chitani olondola opanda kanthu chubu centrifugation ntchito malinga ndi malangizo.

5. Kuipitsa kwina konyansa.

Kusanthula: Zitsanzo zosungidwa kapena zitsanzo zapadera sizimakonzedweratu.

Malangizo: Yang'anirani bwino chitsanzo monga mwalangizidwa.