2 × HiF Taq pamwe neMaster Mix
Katsi Nhamba: HCR2014B
HIF Taq plus Master Musanganiswa (NeDye) ndeye yakagadzirira-kushandisa 2 × premixed mhinduro ine Plus HIF DNA Polymerase, dNTPs, uye yakagadziridzwa buffer.Masoja maviri emonoclonal patembiricha yepakamuri anovharira kuita kwepolymerase uye 3′→5′exonuclease chiitiko anowedzerwa kune master musanganiswa kuti zvive nyore uye zvakanyatsojeka Hot Start PCR.Iyo yekuwedzera chinhu inowedzerwa kune master musanganiswa kupa iyo enzyme kureba chidimbu amplification kugona, kureba kweiyo amplification inogona kusvika ku13 kb, iyo enzyme ine 5'→ 3′ DNA polymerase chiitiko uye 3′→5′. exonuclease chiitiko, kutendeka kwayo kakapetwa makumi masere nenhatu iyo yeTaq DNA polymerase, inova ka9 pane iyo yakajairwa DNA polymerase.Inokodzera kukwidziridzwa kweakaomesesa matemplate, iyo amplification chigadzirwa magumo asina kujeka.
2 × HIF Taq plus Master Mix (NeDye) ine zvakanakira kukurumidza uye nyore, kunzwisiswa kwepamusoro, kusimba kwakasimba, kugadzikana kwakanaka, nezvimwewo, maitiro ekuita anongoda kuwedzera maprimers uye matemplate, uye anogona kukwidziridzwa nembiri- nhanho protocol, kurerutsa matanho ekuyedza uye kuchengetedza nguva.Ichi chigadzirwa chine electrophoresis chiratidzo dhayi, uye PCR zvigadzirwa zvinogona kushandiswa zvakanangana ne electrophoresis.Uye zvakare, chigadzirwa chinewo chaiyo inodzivirira mumiriri, kuitira kuti tenzi musanganiswa ugone kuchengetedza yakagadzikana chiitiko mushure mekudzokorora kutonhora-kunyungudika.
Storage Conditions
Zvigadzirwa zvinofanirwa kuchengetwa pa -25 ~ -15 ℃ kwegore rimwe.
Zvinotsanangurwa
| Product specification | Master Mix |
| Concentration | 2× |
| Sravana Sameeralu Serial 4th Hot Start | Yakavakwa-mukati Hot Start |
| Overhang | Blunt |
| Reaction speed | Rapid |
| Saizi (Chigadzirwa Chekupedzisira) | Kusvika ku13kb |
| Mamiriro ekufambisa | Dry ice |
| Chigadzirwa mhando | High kutendeka PCR premixes |
Mirayiridzo
1.PCR Reaction System
| Zvikamu | Vhoriyamu (μL) |
| DNA Template | Inokodzera |
| Pamberi pekutanga (10 μmol/L) | 2.5 |
| Reverse Primer (10 μmol/L) | 2.5 |
| 2 × HIF Taq pamwe Master Mix | 25 |
| ddH2O | kusvika ku50 |
2.Inokurudzirwa kushandiswa kwematemplate akasiyana
| Rudzi rwe template | Wedzera zvimedu kubva ku1kb kusvika ku10 kb |
| Genomic DNA | 50ng-200 ng |
| Plasmid kana Viral DNA | 10pg-20ng |
| cDNA | 1-2.5 µL (Usapfuure gumi muzana yekupedzisira PCR reaction volume) |
3.Amplification Protocol
1) Matanho maviri Protocol (yakaoma template)
| Cycle step | Temp. | Nguva | Cycles |
| Initial denaturation | 98℃ | 3 min | 1 |
| Denaturation | 98℃ | 10sec | 30-35 |
| Extension | 68℃ | 30 sec/kb | |
| Final extension | 72℃ | 5 min | 1 |
2) Matanho matatu Protocol (yakajairika protocol)
| Cycle step | Temp. | Nguva | Cycles |
| Initial denaturation | 98℃ | 3 min | 1 |
| Denaturation | 98℃ | 10sec | 30-35 |
| Annealing | 60 ℃ | 20 sec | |
| Extension | 72℃ | 30 sec/kb | |
| Final extension | 72℃ | 5 min | 1 |
3) Annealing Gradient Protocol (yakaoma template)
| Cycle step | Tembiricha | Nguva | Cycles |
| Initial denaturation | 98℃ | 3 min | 1 |
| Denaturation | 98℃ | 10 sec | 15 (1 ℃ kuderedza kutenderera) |
| Gradient annealing | 70-55 ℃ | 20 sec | |
| Extension | 72℃ | 30 sec/kb | |
| Denaturation | 98℃ | 10 sec |
20 |
| Annealing | 55℃ | 20 sec | |
| Extension | 72℃ | 30 sec/kb | |
| Final extension | 72℃ | 5 min | 1 |
Zvimiro pasi peakasiyana amplification protocol
| Protocol | Matanho maviri | Nhanho nhatu | Gradient annealing |
| Spec. | fast | pakati | slow |
| Zvakananga | high | pakati | high |
| PCR goho | pakati | high | pakati |
| Detection rate | high | pakati | high |
Notes
Ndokumbira upfeke iyo PPE inodiwa, jasi rakadaro uye magurovhosi, kuve nechokwadi chehutano hwako uye kuchengetedzeka!
