Ka ho toba RT-qPCR Kit II
Tlhaloso ea Kit:
◮Direct RT-qPCR Kit, e sebelisa Foreasy Reverse Transcriptase le Foreasy HS Taq DNA Polymerase e hlahisoang ke Foregene, e nang le buffer e ikhethang ea karabelo, e etsa hore kit ena e be le khanyetso e matla le e tsamaellanang, 'me e ka leka ka kotloloho ts'ebeliso ea sistimi ea Foregene Lysis joalo ka template.Setsi sena se ka etsa kopo bakeng sa lisebelisoa tsa ho bona tsa COVID-19 nucleic acid le nts'etsopele ea lisebelisoa tsa ho lemoha libaktheria tsa pathogenic tsa nucleic acid.
Tlhaloso
Direct RT-qPCR Kit e fana ka buffer e pakiloeng ka thoko ea RT-qPCR, Foreasy HS Taq DNA Polymerase e sebetsang hantle haholo.le Foreasy Reverse Transcriptase(MMLV),e etsa hore e be bonolo bakeng sa ho theha lisebelisoa tsa morao-rao le tokiso ea sistimi, ka har'a netefatso e bonolo.
Direct RT-qPCR Kit, e sebelisa Foreasy Reverse Transcriptase le Foreasy HS Taq DNA Polymerase e hlahisoang ke Foregene, e nang le buffer e ikhethang ea karabelo, e etsa hore kit ena e be le khanyetso e matla le e tsamaellanang, 'me e ka leka ka kotloloho ts'ebeliso ea sistimi ea Foregene Lysis joalo ka template.Setsi sena se ka etsa kopo bakeng sa lisebelisoa tsa ho bona tsa COVID-19 nucleic acid le nts'etsopele ea lisebelisoa tsa ho lemoha libaktheria tsa pathogenic tsa nucleic acid.
Sesebelisoa sena se ka ba karolo ea sehlahisoa sa IVD ka ho toba, se sebelisa habonolo le ka potlako.E hloka feela netefatso e bonolo, ntle le ho ntlafatsoa hape.
Litlhaloso
500T, 50,000T
Likarolo tsa Kit
| Likahare tsa Kit (Sistimi ea karabelo ea 20 μL) | IM-05111-02 | IM-05112-02 |
| 500 T | 50,000 T | |
| Ho bonolo Reverse Transcriptase (200 U/μL) | 50 μL×1 | 1 mL×5 |
| HS Taq DNA Polymerase (5 U/μL) | 100 μL×1 | 1 mL×10 |
| 2 × RT-qPCR e tobilengBuffer | 1 mL × 5 | 500 mL × 1 |
| Taelo | 1 | 1 |
Mehato ea Ts'ebetso
A: Lokisetsa template le reagent
1. Lokisetsa template ea RNA e lokiselitsoeng (Ho kgothaletswa ho sebelisa Foregene Total RNA Isolation Kit series kit ho ntša le ho hloekisa template ea RNA) kapa sehlahisoa sa ho senya sehlahisoa (Ho khothalletsoa ho sebelisa tsamaiso ea Foregene Lysis), li-primers tse khethehileng (10 μM) le lisebelisoa tse ling tse amehang, sesebelisoa.
2. Beha 2 × Direct RT-qPCR Buffer, RNase-Free ddH2O le 20× ROX Reference Dye (haeba ho hlokahala) ka lebokoseng le leqhoa, ho etsa hore tsena li qhibilihe ka tlhaho, 'me li kopane ka bonolo.
B: Lokisetsa tsamaiso ea RT-qPCR
Fumana halofo ea molumo oa buffer ea reaction system (mohlala, haeba bophahamo ba sistimi e le 20 μL, e hloka ho fumana 10 μL 2× Direct RT-qPCR Buffer.). Bakeng sapalo e lekanang ea enzyme,re fana ka maikutlo a horeM-MLV: 10-30 U / 20 μL sistimi ea karabelo,TaqDNA polymerase:1-2 U / 20 μL tsamaiso ea karabeloena ke tlhahiso ea rona, o lokela ho e leka le ho e lokisa), eketsa thempleite ea RNA, li-primers le li-probes, 'me u kenye RNase-Free ddH2O ho isa ho 20 μL.Tokiso e ikhethileng ea sistimi ea karabelo ea RT-qPCR e ka fetisetsoa tafoleng e latelang 1.
Letlapa la 1: Lokisetsa tsamaiso ea RT-qPCR
| Karolo | Bolumo | Khatiso ea ho qetela |
| 2 × Direct RT-qPCR Buffer | 10 μL | 1× |
| Foreasy Reverse Transcriptase(MMLV) | 10-30 U | |
| HS Taq DNA Polymerase | 1-2 U | |
| Primer ea pele(10 μM) | 0.8 μL | 50-900nM |
| Reverse Primer(10 μM) | 0.8 μL | 50-900nM |
| Probe(4 μM) | 1 μL | 200nM |
| Setšoantšo (RNA kapa Lysate) | X μL | |
| 20 × ROX Reference Dye | - | 1* |
| RNase-FreeddH2O | (6.4-X) μL | |
| Kakaretso ea Bolumo | 20 μL |
Tlhokomeliso: Forward Primer le Reverse Primer ke li-primer tse khethehileng tsa lefutso.Sistimi ea qPCR e ka fetoloa ho latela mefuta ea liteko le PCR.Re khothaletsa 400nM bakeng sa khatello ea ho qetela ea li-primers tse ngata.Ka kopo, fetola boholo ba li-primers le li-probes ho latela khatello e lokiselitsoeng le mohopolo oa ho qetela o khothaletsoang.
1 *: Khetha motsoako o loketseng oa ho qetela oa ROX Reference Dye ho latela lisebelisoa tse fapaneng tsa PCR.Khokahano e nepahetseng ea ROX Reference Dye ea lisebelisoa tse tloaelehileng tsa PCR e bonts'oa tafoleng e latelang:
| Lisebelisoa tsa PCR tse ngata | Khatiso ea ho qetela ea ROX Reference Dye |
| ABI PRISM 7000/7300/7700/7900HT/Mohato oa Pele,etc. | 1× (mohlala, eketsa 1μl 20×ROX Reference Dye ho 20 μL reaction system) |
| ABI 7500, 7500 Fast, Stratagene Mx3000P, Mx3005P le Mx4000, joalo-joalo. | 0.5× (mohlala, eketsa 0.5μl 20×ROX Reference Dye ho 20 μL reaction system) |
| Sesebelisoa sa Roche PCR, sesebelisoa sa Bio-Rad PCR, sesebelisoa sa PCR sa Eppendorf, joalo-joalo. | Ntle le ROX Reference Dye |
Storage le Shelf bophelo
Thepa e lokela ho bolokoa ho -20 ± 5 ℃.E lokela ho bolokoa hang-hang ka -20 ℃ sehatsetsing sa thermostatic.Nako ea ho sebetsa ke lilemo tse 2 haeba e bolokiloe boemong bo loketseng.
