Mohato o le Mong ka Potlako RT-qPCR Probe Premix-UNG
Nomoro ea katse: HCR5143A
Mohato o Mong oa RT-qPCR Probe Kit (FOR FAST) ke sesebelisoa sa ho lemoha ka potlako sa RT-qPCR se loketseng single-plex kapa multiplex quantitative PCR se sebelisa RNA joalo ka template (joalo ka vaerase ea RNA).Sehlahisoa sena se sebelisa moloko o mocha oa Taq DNA Polymerase e fetotsoeng ea antibody le mohato o le mong o inehetseng oa Reverse Transcriptase, e nang le buffer e ntlafalitsoeng bakeng sa ho holisa ka potlako, e nang le lebelo la ho holisa ka potlako, bokhoni bo phahameng ba ho holisa le bo ikhethang.E tšehetsa amplification e leka-lekaneng ka bobeli-plex le multiplex ea lisampole tse tlaase le tse phahameng tsa mahloriso ka nako e khutšoanyane.
Likaroloana
1. 5×RT-qPCR buffer (U+)
2. Motsoako oa enzyme (U+)
Lintlha:
a.5×RT-qPCR buffer (U+) e kenyelletsa dNTP le Mg2+;
b.Motsoako oa enzyme (U+) o kenyelletsa reverse transcriptase, Hot Start Taq DNA polymerase, RNase inhibitor le UDG;
c.Sebelisa malebela a Mahala a RNase, li-tubes tsa EP, joalo-joalo.
Pele o sebelisoa, kopanya ka botlalo buffer ea 5×RT-qPCR (U+).Haeba ho na le lipula ka mor'a ho qhibiliha, emela hore buffer e khutlele mocheso oa kamore, kopanya le ho qhibiliha, ebe u li sebelisa ka tloaelo.
Maemo a polokelo
Sehlahisoa se romelloa ka leqhoa le omileng 'me se ka bolokoa ho -25 ~ -15 ℃ bakeng sa selemo sa 1.
Litaelo
1. Boitšoaro Tsamaiso
| Likaroloana | Bophahamo (20 μL karabelo) |
| 2 × RT-qPCR buffer | 4μL |
| Motsoako oa enzyme (U+) | 0.8μL |
| Pele Pele | 0.1~ 1.0μM |
| Primer Reverse | 0.1~ 1.0μM |
| TaqMan Probe | 0.05~0,25μM |
| Setšoantšo | X μL |
| Metsi a se Nang RNase | ho fihlela ho 25μL |
Lintlha: Bophahamo ba karabelo ke 10-50μL.
2. Protocol ea Libaesekele (Stloaelehileng)
| Baesekele mohato | Temp. | Nako | Lipotoloho |
| Reverse Transcription | 55 ℃ | 10 mets | 1 |
| Denaturation ea pele | 95 ℃ | 30 metsotsoana | 1 |
| Denaturation | 95 ℃ | 10 sec | 45 |
| Katoloso/ Katoloso | 60 ℃ | 30 metsotsoana |
| Protocol ea Libaesekele (Ka potlako) Baesekele mohato |
Temp. |
Nako |
Lipotoloho |
| Reverse Transcription | 55 ℃ | 5 mets | 1 |
| Denaturation ea pele | 95 ℃ | 5 s | 1 |
| Denaturation | 95 ℃ | 3 s | 43 |
| Katoloso/ Katoloso | 60 ℃ | 10 s |
Lintlha:
a.Thempereichara ea morao-rao e pakeng tsa 50 ℃ le 60 ℃, ho eketsa mocheso ho thusa ho holisa meaho e rarahaneng le litempele tse phahameng tsa CG;
b.Mocheso o nepahetseng oa annealing o hloka ho lokisoa ho latela boleng ba Tm ba primer, 'me u khethe nako e khuts'oane ea pokello ea matšoao a fluorescence ho latela sesebelisoa sa 'Nete sa Nako ea PCR.
Lintlha
Ka kopo, roala PPE e hlokahalang, jase e joalo ea lab le liatlana, ho netefatsa bophelo bo botle le polokeho ea hau!
