Hoʻokahi ʻanuʻu wikiwiki RT-qPCR Probe Premix-UNG
Helu popoki: HCR5143A
One Step RT-qPCR Probe Kit (FOR FAST) he kit ʻike wikiwiki RT-qPCR i hoʻokumu ʻia i kūpono no ka PCR hoʻokahi-plex a i ʻole multiplex quantitative PCR me ka hoʻohana ʻana i ka RNA ma ke ʻano he maʻi (e like me ka RNA virus).Ke hoʻohana nei kēia huahana i kahi hanauna hou o ka antibody-modified Taq DNA Polymerase a me hoʻokahi ʻanuʻu i hoʻolaʻa ʻia ʻo Reverse Transcriptase, me kahi buffer i hoʻopaʻa ʻia no ka hoʻonui wikiwiki ʻana, ʻoi aku ka wikiwiki o ka amplification, ʻoi aku ka maikaʻi o ka amplification a me ka kikoʻī.Kākoʻo ia i ka hoʻonui ʻana i ka hoʻonui ʻana i ka hoʻokahi-plex a me ka multiplex o nā haʻawina haʻahaʻa a kiʻekiʻe i ka manawa pōkole.
Nā ʻāpana
1. 5×RT-qPCR pale (U+)
2. Huihui Enzyme (U+)
Nā memo:
a.5×RT-qPCR buffer (U+) me dNTP a me Mg2+;
b.ʻO ka hui enzyme (U+) e pili ana i ka reverse transcriptase, Hot Start Taq DNA polymerase, RNase inhibitor a me UDG;
c.E hoʻohana i nā ʻōlelo aʻoaʻo RNase-Free, nā paipu EP, etc.
Ma mua o ka hoʻohana ʻana, e hui pono i ka 5×RT-qPCR buffer (U+).Inā loaʻa ka ua ma hope o ka hoʻoheheʻe ʻana, e kali no ka hoʻi ʻana o ka pahu i ka lumi wela, hui a hoʻoheheʻe ʻia, a laila hoʻohana maʻamau.
Nā Kūlana Waihona
Hoʻouna ʻia ka huahana me ka hau maloʻo a hiki ke mālama ʻia ma -25 ~ -15 ℃ no 1 makahiki.
Nā kuhikuhi
1. Pane Pūnaehana
| Nā ʻāpana | Volume (20 μL pane) |
| 2 ×RT-qPCR pale | 4μL |
| Ka hui ʻana o ka enzyme (U+) | 0.8μL |
| Puhi mua | 0.1~ 1.0μM |
| Kuhikuhi Huli | 0.1~ 1.0μM |
| TaqMan Probe | 0.05~0.25μM |
| Papahana | X μL |
| Ka wai ʻole RNase | a hiki i 25μL |
Nānā: ʻO 10-50μL ka nui o ka pane.
2. Kaʻikala Kaʻa (Standard)
| Kaapuni ʻanuʻu | Temp. | Manawa | Nā pōʻaiapuni |
| Huli Hoʻopaʻa | 55 ℃ | 10 min | 1 |
| Denaturation mua | 95 ℃ | 30 sec | 1 |
| Denaturation | 95 ℃ | 10 sec | 45 |
| Hoʻopili/Hoʻonui | 60 ℃ | 30 sec |
| Kūkākūkā Kaʻikala (wikiwiki) Kaapuni ʻanuʻu |
Temp. |
Manawa |
Nā pōʻaiapuni |
| Huli Hoʻopaʻa | 55 ℃ | 5 min | 1 |
| Denaturation mua | 95 ℃ | 5 s | 1 |
| Denaturation | 95 ℃ | 3 s | 43 |
| Hoʻopili/Hoʻonui | 60 ℃ | 10 s |
Nā memo:
a.ʻO ka mahana transcription hoʻohuli ʻia ma waena o 50 ℃ a 60 ℃, e hoʻonui ana i ka mahana e kōkua i ka hoʻonui ʻana i nā hale paʻakikī a me nā kumu hoʻohālike CG kiʻekiʻe;
b.Pono e hoʻoponopono ʻia ka mahana annealing maikaʻi loa ma muli o ka waiwai Tm o ke kumu mua, a koho i ka manawa pōkole loa no ka hōʻiliʻili ʻana i nā hōʻailona fluorescence ma muli o ka mea kani PCR Real Time.
Nā memo
E ʻoluʻolu e ʻaʻahu i ka PPE pono, e like me ka lole lab a me nā mīkina lima, e hōʻoia i kou olakino a palekana!
