2× HiF Taq oo lagu daray Master Mix

Bisadda No: HCR2014B

Xidhmada: 1ml/5ml/25ml

HIF Taq oo lagu daray Master Mix (With Dye) waa diyaar u ah in la isticmaalo 2× xal premixed oo ka kooban Plus HIF DNA Polymerase, dNTPs, iyo wax hagaajin ah.

Soo dir weydiin

Sharaxaada Alaabta

Faahfaahinta alaabta

Bisadda No: HCR2014B

HIF Taq oo lagu daray Master Mix (With Dye) waa diyaar u ah in la isticmaalo 2× xal premixed oo ka kooban Plus HIF DNA Polymerase, dNTPs, iyo wax hagaajin ah.Laba unugyada difaaca jirka ee monoclonal ee heerkulka qolka ee xannibaya dhaqdhaqaaqa polymerase iyo 3′ 5′exonuclease dhaqdhaqaaqa ayaa lagu daraa isku dhafka sayidka si fudud oo aad u gaar ah PCR Hot Start.Qodobka kordhinta waxaa lagu daraa isku dhafka sayidku si uu u siiyo ensaymka awood dheeraansho jajab ah, dhererka kor u qaadida wuxuu noqon karaa ilaa 13 kb, enzymku wuxuu leeyahay 5'→3' dhaqdhaqaaqa polymerase DNA iyo 3'→5′ Dhaqdhaqaaqa exonuclease, daacadnimadiisu waa 83 jeer taq DNA polymerase, taas oo 9 jeer ka badan polymerase DNA-ga caadiga ah.Waxay ku habboon tahay kor u qaadida moodooyinka adag, badeecada kor u qaadida ayaa ah dhamaadka baraf.

2×HIF Taq oo lagu daray Master Mix (With Dye) waxay leedahay faa'iidooyinka degdegga ah oo sahlan, dareenka sare, gaar ahaan xooggan, xasilloonida wanaagsan, iwm., nidaamka falcelinta kaliya wuxuu u baahan yahay inuu ku daro aasaasayaasha iyo qaababka, waxaana lagu kordhin karaa laba- borotokoolka tillaabada, fududaynta tillaabooyinka tijaabada ah iyo badbaadinta wakhtiga.Alaabtani waxa ay ka kooban tahay dheeha tusaha electrophoresis, iyo alaabta PCR waxa si toos ah loogu isticmaali karaa electrophoresis.Intaa waxaa dheer, alaabtu waxay sidoo kale ka kooban tahay wakiil gaar ah oo ilaalin ah, si isku-darka sayidku u sii wadi karo dhaqdhaqaaqa xasilloon ka dib marka la qaboojiyo-dhalasho soo noqnoqda.

Hore: 2×PCR Master Mix (la'aanteed) HCR2013B

Xiga: Qalabka PCR ee tooska ah ee dhirta HCR2020A

Xaaladaha Kaydinta

Alaabta waa in lagu kaydiyaa at -25 ~ -15 ℃ 1 sano.

Tilmaamaha

Tilmaamaha alaabta	Master Mix
Xoog-saarid	2×
Bilow kulul	Ku-dhismay Hot Start
Dulqaadasho	Maqan
Xawaaraha falcelinta	Degdeg ah
Baaxadda (Badeecada u dambaysa)	Ilaa 13kb
Shuruudaha gaadiidka	Baraf qallalan
Nooca alaabta	Kalsoonida sare ee PCR premixes

Tilmaamaha

1.Nidaamka falcelinta PCR

Qaybaha	Mugga (μL)
Qaabka DNA-da	Ku haboon
Qoryaha hore ee hore (10 μmol/L)	2.5
Dib-u-celinta hore (10 μmol/L)	2.5
2×HIF Taq oo lagu daray Master Mix	25
ddH₂O	ilaa 50

2.Lagu taliyay isticmaalka habab kala duwan

Nooca template	Kordhi jajabyada 1kb ilaa 10kb
DNA-da genomiga ah	50ng-200 ng
Plasmid ama Viral DNA	10pg-20ng
CDNA	1-2.5 µL (Ha dhaafin 10% mugga falcelinta PCR ee u dambeeya)

3.Kordhinta Hab-maamuuska

1) Hab-maamuuska Laba-Tallaabo (qaabka kakanaanta)

Tallaabo wareeg	Heerkulka	Waqtiga	Wareegtada
Denaturation bilowga ah	98℃	3 daq	1
Denaturation	98℃	10 ilbiriqsi	30-35
Kordhinta	68℃	30 ilbiriqsi/kb	30-35
Kordhinta kama dambaysta ah	72℃	5 daqiiqo	1

2) Hab-maamuuska Saddex-tilaabo (Borotokoolka caadiga ah)

Tallaabo wareeg	Heerkulka	Waqtiga	Wareegtada
Denaturation bilowga ah	98℃	3 daq	1
Denaturation	98℃	10 ilbiriqsi	30-35
Annealing	60℃	20 ilbiriqsi
Kordhinta	72℃	30 ilbiriqsi/kb
Kordhinta kama dambaysta ah	72℃	5 daqiiqo	1

3) Hab-maamuuska Gradient Annealing (qaabka kakanaanta)

Tallaabo wareeg	Heerkulka	Waqtiga	Wareegtada
Denaturation bilowga ah	98℃	3 daq	1
Denaturation	98℃	10 ilbiriqsi	15 (1℃ dhimis halkii wareeg)
Ciribtirka tartiib tartiib ah	70-55 ℃	20 ilbiriqsi
Kordhinta	72℃	30 ilbiriqsi/kb
Denaturation	98℃	10 ilbiriqsi	20
Annealing	55℃	20 ilbiriqsi
Kordhinta	72℃	30 ilbiriqsi/kb
Kordhinta kama dambaysta ah	72℃	5 daqiiqo	1

Astaamaha hoos yimaada borotokoolka kordhinta ee kala duwan

Protocol	Labo-Tallaabo	Saddex tallaabo	Ciribtirka tartiib tartiib ah
Spec.	degdeg ah	dhexdhexaad ah	gaabis ah
Gaar ahaan	sare	dhexdhexaad ah	sare
Wax-soo-saarka PCR	dhexdhexaad ah	sare	dhexdhexaad ah
Heerka ogaanshaha	sare	dhexdhexaad ah	sare