Falanqaynta soo socota ee dhibaatooyinka suurtagalka ah ee gudahaBuccalsuuf/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.

Tiirka sifaynta ayaa xidhan

Qalabkan, hawlgalka soo saarista DNA-da genomic, tiirka sifaynta ayaa si toos ah loogu dhejiyaa muunadda lysis ee isku-darka enzymatic iyada oo aan la qaadin tallaabo centrifugation, iyo tiirka nadiifinta ayaa laga yaabaa in la xannibo sababtoo ah enzymization aan dhamaystirnayn iyo viscosity sare ee muunadda.

Sababaha suurtagalka ah ee soo socda waa sida soo socota:

1. Dheefshiidka enzymatic oo aan dhamaystirnayn ee shaybaarada unugyada.

Talo-soo-jeedin: Waqtiga muunad-samaynta ee Foregene Protease si habboon ayaa loo kordhin karaa ama ka sarreeya ayaa la qaadi karaa ka dib marka la isku daro 12,000 rpm (~ 13,400× g) 5 daqiiqo.

2. Isticmaalka xad dhaafka ah ee shaybaarada unugyada ama unugyo waaweyn.

Talo soo jeedin: Way fiicantahay inaadan dhaafin 1Buccal suufka saamiga;haddii muunadku aad u weyn yahay, kordhi qiyaasta Buffer ST1, Foregene Protease, kaydinta ST2 si waafaqsan.

3. Mawduuca muunada oo aad u sarreeya.

Talo soo jeedin: Muundooyinka waxaa si habboon loogu qasi karaa 10 mM ee Tris-HCl ka hor inta aan la soo saarin DNA-da genomic.

4. Jajabka kaadhka dhiigga oo la nuugay.

Talo soo jeedin: Waqtiga dhexe ee ku meel gaadhka ah ee tallaabada 6 ee dhibicda dhiigga (FTA Card) soo saarista genomic ayaa si habboon loo kordhin karaa.

Waxsoosaar hooseeya ama DNA-da oo aan lahayn

Inta badan waxaa jira arrimo kala duwan oo saameeya dhalidda DNA-ga, oo ay ku jiraan asalka muunada, xaaladaha kaydinta muunada, diyaarinta muunada, wax-is-dabamarinta, iwm.

DNA-da genomic lama heli karo marka la soo saarayo

Sababaha suurtagalka ah waa sida soo socota:

1. Ilaalinta aan habboonayn ee shaybaarada ama kaydinta wakhti dheer waxay keenaysaa hoos u dhaca DNA-da.

Talo soo jeedin: Suugaanta afka waa in la door bidaa in si cusub loo muunadeeyo, lagumana talinayo in la isticmaalo suufyada la ilaaliyo ee hawlaha soo saarista DNA-da;shaybaarrada dhibcaha dhiigga waa in ay xaqiijiyaan in tayadu ay tahay mid u qalanta oo wakhtiga kaydinta aanu noqon mid aad u dheer.

2. Isticmaalka nudaha oo aad u yar waxay keeni kartaa in aan la soo saarin DNA-da genomic ee u dhiganta.

Talo soo jeedin: Raacbuccal tilmaan-bixinta muunadda ee hagaha hawlgalka, oo tirtir inta jeer ee suurtogalka ah si unugyo ku filan loogu dhejiyo suufka afka si loo soo saaro DNA-da genomic;Soo saarista saamiga dhiigga, meesha gooynta dhibicda dhiigga ayaa si habboon loo kordhin karaa.

3. Protease Foregene si aan munaasib ahayn ayaa loo ilaaliyaa, taasoo keenta hoos u dhac ku yimaada dhaqdhaqaaqiisa ama hawlgelin la'aan.

Talo soo jeedin: Xaqiiji xaaladaha kaydintaFOregene Protease ama ku beddel Foregene Protease cusub ee falcelinta enzymatic.

4. Ilaalinta aan saxda ahayn ee xirmada ama wakhtiga kaydinta ayaa aad u dheer, taasoo keentay fashilka qaybaha qaar ee xirmada.

Talo soo jeedin: Iibso mid cusubBuccal DNA dhuubankarantiil xirmada hababka la xiriira.

5. Buffer WB kuma darto ethanol dhammaystiran.

Talo soo jeedin: Xaqiiji in kaydiyaha WB uu ku darayo mugga saxda ah ee ethanol dhammaystiran.

6. Eluent si sax ah looguma darin filimka silikoon.

Talo soo jeedin: Ku dar 65°CEluent horay loo diiray ayaa hoos ugu dhacda bartamaha xuubka silikoonka oo ka tag heerkulka qolka 5 min si loo kordhiyo waxtarka sare.

LDNA-da genomic-soo-saarka go'doonsan

Sababaha suurtagalka ah ee soo socda waa sida soo socota:

1. Ilaalinta aan habboonayn ee shaybaarada ama kaydinta wakhti dheer waxay keenaysaa hoos u dhaca DNA-da.

Talo-soo-jeedin: Suuxdin afka laga qaato ayaa la door bidaa in mar cusub laga qaaday muunad, suufka la xafidayna waa inaan loo isticmaalin soo saarista DNA-da genomic.

2. Haddii xadiga muunada unuggu aad u yar yahay, waxa ku jira DNA-ga genomiga ah ee la soo saaray wuu yaraan doonaa.

Talo soo jeedin: Raac tilmaamaha muunadda suufka afka ee ku jira hagaha qalliinka, adigoo masaxaya inta jeer ee suurtogalka ah si unugyo ku filan loogu dhejiyo suufka afka laga soo saaro DNA-da genomic.

3. Protease Foregene si aan munaasib ahayn ayaa loo ilaaliyaa, taasoo keenta hoos u dhac ku yimaada dhaqdhaqaaqiisa ama hawlgelin la'aan.

Talo soo jeedin: Xaqiiji xaaladaha kaydintathe Foregene Protease ama ku beddel mid cusub Foregene Protease ee falcelinta enzymatic.

4. Dhibaatooyin aan caadi ahayn.

Talo soo jeedin: Isticmaal Buffer EB si aad u iftiimiso;haddii aad isticmaalayso ddH₂O ama kuwa kale, xaqiiji in pH ee eluate-ku uu u dhaxeeyo 7.0-8.5.

5. Eluate si sax ah looguma darin hoos u dhaca.

Talo soo jeedin: Ku dar dhibco eluent badhtamaha xuubka silikoonka oo ku dhaaf heerkulka qolka 5 min si loo kordhiyo waxtarka elution.

6. Dareeraha elation ayaa urura wax aad u yar.

Talo-soo-jeedin: U isticmaal eluent ee elution DNA genomic sida loogu baahan yahay tilmaamaha, ugu yaraanaan ka yarayn in ka badan 15μl.

Low daahirnimo of DNA-da genomicgo'doonsan

Nadiifinta DNA-da ee hooseeya waxay u horseedi kartaa guuldarro ama natiijooyin aan ku qanacsanayn tijaabooyinka hoose, sida: enzymes lama goyn karo, PCR ma heli karto jajabka hidda-socodka xiisaha, iwm.

Sababaha suurtagalka ah waa sida soo socota:

1. Wasakhowga Heteroprotein, wasakhowga RNA.

Falanqaynta: Tiirka sifaynta laguma maydhin iyadoo la isticmaalayo Buffer PW;tiirka nadiifinta dhaqidda Buffer PW laguma dhaqin iyadoo la isticmaalayo xawaaraha dhexe ee saxda ah.

Talo-soo-jeedin: Hubi in aanay jirin wax roob ah ka hor inta aan lagu darin ethanol;Iska hubi inaad dhaqdo tiirka nadiifinta si waafaqsan tilmaamaha, tallaabadanna lama dhaafi karo.

2. wasakhowga wasakhda wasakhaysan.

Falanqaynta: Buffer WB tiirka nadiifinta ayaa laga saaray ama la dhaqay hal mar oo kaliya, taasoo keentay faddarayn ionic ah.

Talo soo jeedin: Hubi inaad dhaqdo Buffer WB 2 jeer sida lagu faray si aad meesha uga saarto ions haraaga intii suurtogal ah.

3. RNA enzyme wasakhaysan.

Falanqaynta: RNases shisheeye ayaa lagu daray kayd;Qalliinka dhaqidda Buffer PW sax ma ahayn, taasoo keentay hadhaaga RNase, saamaynaya hawlaha tijaabada ah ee RNA-da hoose, sida qoraalka in vitro.

Talo soo jeedin: Aashitada nukleic ee taxanaha ahisolaXirmooyinka qalabka ayaa ka saari kara RNA iyada oo aan lagu darin RNase dheeraad ah,sidaas Qalabka Go'doominta DNA-ga ee Swab/FTAloo baahan yahay't ku dar RNase;Hubi inaad raacdo tilmaanta Buffer PW tiirka nadiifinta, tallaabadan lama dhaafi karo.

4. Haraaga ethanol.

Falanqaynta: Buffer WB ma samayn centrifugation tube madhan ka dib markii la dhaqay tiirka sifaynta.

Talo soo jeedin: Samee hawlgalka centrifugation tube madhan saxda ah sida ku xusan tilmaamaha.

5. wasakhowga kale ee wasakhaysan.

Falanqaynta: Muunado kaydsan ama muunado gaar ah lama sii daawayn.

Talo soo jeedin: Si fiican ugu sii daawee muunada sida lagu faray.