RT-LAMP Colormetric Master Mix HCB5204A
Le mveliso iqulethe isithinteli reaction, RT-Enzymes Mix (Bst DNA polymerase kunye nobushushu-resistant reverse transcriptase), lyophilized Protectants kunye amacandelo chromogenic idayi.Ukusebenzisa, sebenzisa nje i-Buffer, i-enzyme yokusabela kunye ne-primer ixutywe kwaye yongezwa kwi-template;ukongeza umkhuseli lyophilized kunokuba ngqo.Yayixhunywe kwi-lyophilizer kunye ne-lyophilized, kwaye kuphela i-primers kunye neetemplates zongezwa xa zisetyenziswa.Le khithi ibonelela ngokukhawuleza, ukukhangela okubonakalayo okucacileyo kokukhulisa, ukusabela okungalunganga kubonakaliswe kubomvu kunye nokusabela okulungileyo kuboniswa ngotshintsho oluphuzi.
Icandelo
| Icandelo | HCB5204A-01 | HCB5204A-02 | HCB5204A-03 |
| Isithinteli soKwandiswa kwe-Loop-mediated (ngedayi) | 0.96 mL | 4.80 mL×2 | 9.60 mL×10 |
| I-RT-Enzymes Mix | 270 μL | 2.70 mL | 2.70 mL×10 |
| Umkhuseli we-Lyophilized | 0.96 mL×2 | 9.60 ml×2 | 9.60 mL×20 |
Usetyenziso
YeDNA okanye iRNA isothermal amplification.
Iimeko zokuGcina
Ithuthwa ngoMkhenkce owomileyo, ogcinwe ku -25~ -15℃.Gwema ukunyibilika rhoqo komkhenkce, imveliso isebenza kwiinyanga ezili-12.
Umgaqo-nkqubo
1.Tyibilika isithinteli sokusabela esiza kusetyenziswa kwiqondo lobushushu begumbi.I-Vortex ngokufutshane okanye ukuguqula iityhubhu amaxesha amaninzi ukuze ixube ngokucokisekileyo, emva koko i-centrifuge iqokelele ulwelo ukuya ezantsi kumbhobho.
2.Ukulungiswa kwenkqubo yokusabela.Le reagent inokulungiswa kwiinkqubo ezimbini reaction, ulwelo reaction mix kunye lyophilized inkqubo mix.
1) Lungisa umxube wokuphendula ulwelo
| Icandelo | Umthamo |
| Isithinteli soKwandiswa kwe-Loop-mediated (ngedayi) | 10 μL |
| I-RT-Enzymes Mix | 2.8 μL |
| 10 × I-Primer Mixa | 5 μL |
| Izakhelo zeDNA/RNA b | × μL |
| Amanzi angenaNyukliya | Ukuya kuthi ga kwi-50 μL |
2) Inkqubo ye-Lyophilization mix
① Lungisa umxube we-lyophilized
| Icandelo | Umthamo |
| Isithinteli soKwandiswa kwe-Loop-mediated (ngedayi) | 10 μL |
| Umkhuseli we-Lyophilized | 20 μL |
| I-RT-Enzymes Mix | 2.8 μL |
| Amanzi angenaNyukliya | Ukuya kuthi ga kwi-50 μL |
② I-Lyophilization: I-Mix elungiselelwe yayi-lyophilized kwinkqubo ye-50μL
③ Lungisa umxube wokusabela
| Icandelo | Umthamo |
| Umxube we-Lyophilized | Iqhekeza eli-1 |
| 10 × I-Primer Mixa | 5 μL |
| Izakhelo zeDNA/RNA b | × μL |
| Amanzi angenaNyukliya | Ukuya kuthi ga kwi-50 μL |
Amanqaku:
1) a.I-10 × I-Primer Mix : 16 μM FIP / BIP, 2 μM F3 / B3, 4 μM Loop F / B;
2) b.I-DEPC (i-soluble yamanzi) iyacetyiswa kwi-nucleic acid templ.
1.Fukamisa kwi-65 ° C kwi-30-45mins, enokwandiswa ngokufanelekileyo ngokutshintsha umbala Ixesha lokuphendula.
2.Ngokweliso lenyama, umthubi u-positive kwaye ubomvu bu-negative.
Amanqaku
1.Ityuwa inokuvela emazantsi etyhubhu ye-buffer, i-vortex ngokufutshane okanye iguqule iityhubhu amaxesha amaninzi ukuze ixutywe ngokucokisekileyo kwiqondo lobushushu begumbi.
2.Ubushushu bokusabela bunokwenziwa phakathi kwe-62 ℃ kunye ne-68 ℃ ngokwemeko yeeprimers.
3.Ii-reagents ezipakishweyo akufanele zibekwe emoyeni ixesha elide.
4.Ukuguquka kombala obomvu notyheli kuxhomekeke kutshintsho lwe-pH lwenkqubo yokusabela, nceda ungasebenzisi isisombululo sogcino se-Tris nucleic acid, ekucetyiswa ukuba usebenzise i-ddH.2O i-nucleic acid egciniweyo;
5.Uvavanyo kufuneka lubekwe emgangathweni, kubandakanywa ukulungiswa kwenkqubo yokusabela, i-lyophilization, kunye nokucubungula isampuli kunye nenkqubo yokongeza isampuli;
6.Ukuze ugweme ukungcola, kucetyiswa ukuba ulungiselele inkqubo yokusabela kwibhentshi ecocekileyo kakhulu, kwenye Yongeza iitemplates kwi-fume hood yegumbi ukuze ugweme ukuphazamiseka okungalunganga.
