Intambwe imwe Byihuse RT-qPCR Probe Premix-UNG
Injangwe No: HCR5143A
Intambwe imwe RT-qPCR Probe Kit (KUBERA VUBA) nigikoresho gishingiye kuri RT-qPCR cyihuta cyerekana ibikoresho bikwiranye na PCR imwe cyangwa plexlex numero PCR ikoresha RNA nkicyitegererezo (nka virusi ya RNA).Iki gicuruzwa gikoresha igisekuru gishya cya antibody yahinduwe na Taq ADN Polymerase hamwe nintambwe imwe yihariye ya Reverse Transcriptase, hamwe na buffer nziza kugirango yongerwe vuba, ifite umuvuduko mwinshi wo kwihuta, gukora neza kandi neza.Ifasha kuringaniza amplifisione haba muri plex imwe na multiplex ya sample nkeya kandi yibanze cyane mugihe gito.
Ibigize
1. 5 × RT-qPCR buffer (U +)
2. Enzyme ivanze (U +)
Inyandiko:
a.5 × RT-qPCR buffer (U +) ikubiyemo dNTP na Mg2+;
b.Enzyme ivanga (U +) ikubiyemo transcript transcript, Hot Start Taq ADN polymerase, inhibitor ya RNase na UDG;
c.Koresha inama zubusa za RNase, EP tubes, nibindi
Mbere yo gukoresha, vanga neza 5 buffer ya RT-qPCR (U +).Niba hari imvura iguye nyuma yo gushonga, tegereza ko buffer isubira mubushyuhe bwicyumba, vanga kandi ushonga, hanyuma ubikoreshe bisanzwe.
Ububiko
Ibicuruzwa byoherejwe hamwe na barafu yumye kandi birashobora kubikwa kuri -25 ~ -15 ℃ kumwaka 1.
Amabwiriza
1. Igisubizo Sisitemu
| Ibigize | Umubumbe (20 μL reaction) |
| 2 buffer RT-qPCR | 4μL |
| Enzyme ivanga (U +) | 0.8μL |
| Imbere | 0.1 ~ 1.0μM |
| Gusubira inyuma | 0.1 ~ 1.0μM |
| TaqMan | 0.05 ~ 0.25μM |
| Inyandikorugero | X μL |
| Amazi Yubusa | kugeza kuri 25μL |
Icyitonderwa: Ingano yimikorere ni 10-50μL.
2. Amarushanwa yo gusiganwa ku magare (S.tandard)
| Ukuzenguruka intambwe | Ubushuhe. | Igihe | Amagare |
| Guhindura Inyandiko | 55 ℃ | Imin. 10 | 1 |
| Gutandukana kwambere | 95 ℃ | 30 amasegonda | 1 |
| Gutandukana | 95 ℃ | Amasegonda 10 | 45 |
| Kwiyongera / Kwagura | 60 ℃ | 30 amasegonda |
| Umukino wo gusiganwa ku magare (Byihuse) Ukuzenguruka intambwe |
Ubushuhe. |
Igihe |
Amagare |
| Guhindura Inyandiko | 55 ℃ | Imin | 1 |
| Gutandukana kwambere | 95 ℃ | 5 s | 1 |
| Gutandukana | 95 ℃ | 3 s | 43 |
| Kwiyongera / Kwagura | 60 ℃ | 10 s |
Inyandiko:
a.Ubushyuhe bwo kwandukura buri hagati ya 50 ℃ kugeza kuri 60 ℃, kongera ubushyuhe bifasha kwagura imiterere igoye hamwe na CG yuzuye inyandikorugero;
b.Ubushyuhe bwiza bwa annealing bugomba guhinduka ukurikije agaciro ka Tm ya primer, hanyuma ugahitamo igihe gito cyo gukusanya ibimenyetso bya fluorescence ukurikije igikoresho nyacyo PCR.
Inyandiko
Nyamuneka nyamuneka wambare PPE ikenewe, ikote rya laboratoire hamwe na gants, kugirango ubuzima bwawe n'umutekano wawe!
