RT-LAMP Colormetric Master Mix HCB5204A
Mankhwalawa ali ndi chitetezo chamthupi, RT-Enzymes Mix (Bst DNA polymerase ndi reverse transcriptase yosamva kutentha), Ma Protectants a lyophilized ndi zigawo za utoto wa chromogenic.Kuti mugwiritse ntchito, ingogwiritsani ntchito Buffer, ma enzyme ndi zoyambira zimasakanizidwa ndikuwonjezeredwa ku template;Kuonjezera chitetezo cha lyophilized kungakhale kolunjika.Zinalumikizidwa ndi lyophilizer ndi lyophilized, ndipo zoyambira zokha ndi ma templates zidawonjezeredwa zikagwiritsidwa ntchito.Chida ichi chimapereka chidziwitso chofulumira, chowonekera bwino cha kukulitsa, komwe kumapangitsa kuti kukhale koyipa kumasonyezedwa muzofiira komanso zabwino zimasonyezedwa ndi kusintha kwachikasu.
Chigawo
| Chigawo | HCB5204A-01 | HCB5204A-02 | HCB5204A-03 |
| Loop-mediated Amplification Buffer (yokhala ndi utoto) | 0.96 mL | 4.80mL × 2 | 9.60mL × 10 |
| RT-Enzymes Mix | 270 μl | 2.70 ml | 2.70mL × 10 |
| Chitetezo cha Lyophilized | 0.96 mL × 2 | 9.60mL × 2 | 9.60mL × 20 |
Mapulogalamu
Kwa DNA kapena RNA isothermal amplification.
Zosungirako
Kunyamulidwa ndi Dry ice, yosungidwa pa -25 ~ -15 ℃.Pewani kuzizira pafupipafupi, mankhwalawa amakhala kwa miyezi 12.
Ndondomeko
1.Thawtsani chotchinga chomwe chingagwiritsidwe ntchito kutentha kwa firiji.Vortex mwachidule kapena kutembenuza machubu kangapo kuti asakanize bwino, kenako centrifuge kusonkhanitsa madzi pansi pa chubu.
2.Kukonzekera anachita dongosolo.Izi reagent akhoza kukonzekera awiri anachita kachitidwe, madzi anachita kusakaniza ndi lyophilized dongosolo mix.
1) Konzani zosakaniza zamadzimadzi
| Chigawo | Voliyumu |
| Loop-mediated Amplification Buffer (yokhala ndi utoto) | 10 μl pa |
| RT-Enzymes Mix | 2.8 μl |
| 10 × Primer Mixa | 5 ml |
| Zithunzi za DNA / RNA b | × μl ndi |
| Madzi Opanda Nuclease | Mpaka 50 μL |
2) Kusakaniza kwa Lyophilization system
① Konzani kusakaniza kwa lyophilized
| Chigawo | Voliyumu |
| Loop-mediated Amplification Buffer (yokhala ndi utoto) | 10 μl pa |
| Chitetezo cha Lyophilized | 20 μl pa |
| RT-Enzymes Mix | 2.8 μl |
| Madzi Opanda Nuclease | Mpaka 50 μL |
② Lyophilization: Kusakaniza kokonzekera kunali lyophilized mu dongosolo la 50μL
③ Konzani zosakaniza
| Chigawo | Voliyumu |
| Kusakaniza kwa Lyophilized | 1 chidutswa |
| 10 × Primer Mixa | 5 ml |
| Zithunzi za DNA / RNA b | × μl ndi |
| Madzi Opanda Nuclease | Mpaka 50 μL |
Ndemanga:
1) a.10 × Primer Mix : 16 μM FIP / BIP, 2 μM F3 / B3, 4 μM Loop F / B;
2) b.DEPC (madzi osungunuka) amalimbikitsidwa pa nucleic acid templ.
1.Yalirani pa 65 ° C kwa 30-45mins, yomwe imatha kukulitsidwa moyenera molingana ndi kusintha kwa mtundu Nthawi yochitira.
2.Malinga ndi maso, chikasu chinali chabwino ndipo chofiira chinali choyipa.
Zolemba
1.Mchere ukhoza kuwonekera pansi pa chubu chotchinga, vortex mwachidule kapena kutembenuza machubu kangapo kuti usakanize bwino kutentha kwa chipinda.
2.Kutentha komweko kumatha kukonzedwa pakati pa 62 ℃ ndi 68 ℃ malinga ndi momwe ma primers amakhalira.
3.The mmatumba reagents sayenera poyera mpweya kwa nthawi yaitali.
4.Maonekedwe ofiira ndi achikasu amadalira kusintha kwa pH kwa machitidwe, chonde musagwiritse ntchito njira yosungiramo ya Tris nucleic acid, yomwe ikulimbikitsidwa kugwiritsa ntchito ddH.2O kusungidwa nucleic asidi;
5.The kuyesera ayenera standardized, kuphatikizapo yokonza anachita dongosolo, lyophilization, ndi chitsanzo processing ndi chitsanzo kuwonjezera ndondomeko;
6.Kuti mupewe kuipitsidwa, tikulimbikitsidwa kukonzekera kachitidwe kachitidwe mu benchi yoyera kwambiri, mu zina Onjezani ma tempuleti ku chiwombankhanga chachipindacho kuti mupewe kusokonezedwa kwabodza.
