RT-LAMP Colormetric Master Mix HCB5204A
Aia i loko o kēia huahana ka mea ho'ōla ho'ōla, RT-Enzymes Mix (Bst DNA polymerase and heat-resistant reverse transcriptase), lyophilized Protectants a me nā ʻāpana dye chromogenic.No ka hoʻohana, e hoʻohana wale i ka Buffer, ua hui pū ʻia ka enzyme reaction a me ka primer a hoʻohui ʻia i ka template;ʻO ka hoʻohui ʻana i ka pale lyophilized hiki ke pololei.Ua hoʻohui ʻia i kahi lyophilizer a lyophilized, a ua hoʻohui ʻia nā primers a me nā templates i ka wā i hoʻohana ʻia.Hāʻawi kēia pahu i kahi ʻike wikiwiki a maopopo o ka amplification, ʻo ia ka hopena maikaʻi ʻole i hōʻike ʻia i ka ʻulaʻula a hōʻike ʻia ka hopena maikaʻi e ka hoʻololi ʻana i ka melemele.
ʻāpana
| ʻāpana | HCB5204A-01 | HCB5204A-02 | HCB5204A-03 |
| Hoʻopaʻa hoʻopaʻa hoʻonui i hoʻopaʻa ʻia i ka Loop (me ka waihoʻoluʻu) | 0.96 mL | 4.80 mL×2 | 9.60 mL×10 |
| Huihui RT-Enzymes | 270 μL | 2.70 mL | 2.70 mL×10 |
| Lyophilized pale | 0.96 mL×2 | 9.60 mL×2 | 9.60 mL×20 |
Nā noi
No ka DNA a i ʻole RNA isothermal amplification.
Nā Kūlana Waihona
Lawe ʻia me ka hau maloʻo, mālama ʻia ma -25~ -15 ℃.E hōʻalo i ka hoʻoheheʻe pinepine ʻana, pono ka huahana no 12 mahina.
Kūkākūkā
1.E hoʻoheheʻe i ka pale pale e hoʻohana ʻia ma ka wela lumi.Vortex pōkole a hoʻohuli paha i nā paipu i nā manawa he nui e hui pono, a laila centrifuge e hōʻiliʻili i ka wai i lalo o ka paipu.
2.Hoʻomākaukau i ka ʻōnaehana pane.Hiki ke hoʻomākaukau ʻia kēia reagent i ʻelua ʻōnaehana hopena, ka hui ʻana o ka wai a me ka ʻōnaehana lyophilized.
1) E hoʻomākaukau i ka hui ʻana o ka wai
| ʻāpana | Volume |
| Hoʻopaʻa hoʻopaʻa hoʻonui i hoʻopaʻa ʻia i ka Loop (me ka waihoʻoluʻu) | 10 μL |
| Huihui RT-Enzymes | 2.8 μL |
| 10 × Hui Pūmuaa | 5 μL |
| Nā templates DNA/RNA b | × μL |
| Wai Nuclease-free | A hiki i 50 μL |
2) Hui pūnaewele Lyophilization
① E hoʻomākaukau i ka hui lyophilized
| ʻāpana | Volume |
| Hoʻopaʻa hoʻopaʻa hoʻonui i hoʻopaʻa ʻia i ka Loop (me ka waihoʻoluʻu) | 10 μL |
| Lyophilized pale | 20 μL |
| Huihui RT-Enzymes | 2.8 μL |
| Wai Nuclease-free | A hiki i 50 μL |
② Lyophilization: Ua lyophilized ka Mix i hoʻomākaukau ʻia ma kahi ʻōnaehana 50μL
③ E hoʻomākaukau i ka hui ʻana
| ʻāpana | Volume |
| Hoʻohui lyophilized | 1 apana |
| 10 × Hui Pūmuaa | 5 μL |
| Nā templates DNA/RNA b | × μL |
| Wai Nuclease-free | A hiki i 50 μL |
Nā memo:
1) a.10×Primer Mix : 16 μM FIP/BIP, 2 μM F3/B3, 4 μM Loop F/B;
2) b.Paipai ʻia ʻo DEPC (water soluble) no ka templ acid nucleic.
1.Incubate ma 65 ° C no 30-45mins, hiki ke hoʻolōʻihi ʻia e like me ka hoʻololi kala ʻana.
2.Wahi a ka maka ʻōlohelohe, maikaʻi ka melemele a maikaʻi ʻole ka ʻulaʻula.
Nā memo
1.Hiki ke ʻike ʻia ka paʻakai ma lalo o ka paipu paʻa, vortex pōkole a i ʻole e hoʻohuli i nā paipu i nā manawa he nui e hui pono i ka lumi wela.
2.Hiki ke hoʻonui ʻia ka mahana hopena ma waena o 62 ℃ a me 68 ℃ e like me ke ʻano o nā primers.
3.ʻAʻole pono e ʻike ʻia nā reagents i hoʻopaʻa ʻia i ka ea no ka manawa lōʻihi.
4.ʻO ka hopena o ka ʻulaʻula a me ka melemele e pili ana i ka hoʻololi ʻana o ka pH o ka ʻōnaehana pane, e ʻoluʻolu mai hoʻohana i ka Tris nucleic acid storage solution, ʻōlelo ʻia e hoʻohana i ka ddH2ʻO ka waikawa nucleic i mālama ʻia;
5.Pono e hoʻohālikelike ʻia ka hoʻokolohua, me ka hoʻomākaukau ʻana i ka ʻōnaehana pane, lyophilization, a me ka hoʻoheheʻe ʻana a me ke kaʻina hana hoʻohui;
6.No ka pale ʻana i ka haumia, pono e hoʻomākaukau i ka ʻōnaehana pane i loko o kahi papa hoʻomaʻemaʻe ultra-maʻemaʻe, ma nā ʻano hoʻohālikelike ʻē aʻe i ka puʻupuʻu fume o ka lumi e pale aku ai i ka hoʻopili pololei ʻana.
