Inyathelo elinye ngokukhawuleza RT-qPCR Probe Premix-UNG
Inombolo yekati: HCR5143A
Inyathelo elinye RT-qPCR Probe Kit (FOR FAST) yiprobe-based RT-qPCR ikiti yokufumanisa ngokukhawuleza ilungele i-single-plex okanye i-multiplex quantitative PCR isebenzisa i-RNA njenge template (efana nentsholongwane ye-RNA).Le mveliso isebenzisa isizukulwana esitsha se-antibody-modified Taq DNA Polymerase kunye nenyathelo elinye elizinikeleyo i-Reverse Transcriptase, ene-buffer ephuculweyo yokukhulisa ngokukhawuleza, enesantya sokukhulisa ngokukhawuleza, ukusebenza kakuhle kokukhulisa kunye neenkcukacha.Ixhasa ukukhuliswa okulinganayo kuzo zombini i-plex kunye ne-multiplex yeesampuli eziphantsi kunye eziphezulu zoxinaniso ngexesha elifutshane.
Amacandelo
1. 5×RT-qPCR isithinteli (U+)
2. Umxube we-enzyme (U+)
Amanqaku:
a.I-5×RT-qPCR buffer (U+) iquka i-dNTP kunye ne-Mg2+;
b.Umxube we-enzyme (U +) uquka i-reverse transcriptase, i-Hot Start Taq DNA polymerase, i-RNase inhibitor kunye ne-UDG;
c.Sebenzisa iingcebiso ze-RNase-Free, iityhubhu ze-EP, njl.
Ngaphambi kokusetyenziswa, xuba ngokucokisekileyo i-5×RT-qPCR buffer (U+).Ukuba kukho naziphi na iimvula emva kokunyibilika, linda isithinteli ukuba sibuyele kwiqondo lobushushu begumbi, xuba kwaye unyibilike, emva koko uzisebenzise ngokwesiqhelo.
Iimeko zokuGcina
Imveliso ithunyelwa ngeqhwa elomileyo kwaye ingagcinwa kwi -25 ~ -15 ℃ ngonyaka we-1.
Imiyalelo
1. Ukusabela Inkqubo
| Amacandelo | Umthamo (20 μL impendulo) |
| I-2 × RT-qPCR buffer | 4μL |
| Umxube we-enzyme (U+) | 0.8μL |
| I-Primer Phambili | 0.1 ~ 1.0μM |
| I-Primer Reverse | 0.1 ~ 1.0μM |
| TaqMan Probe | 0.05~0,25μM |
| Isifanekiso | X μL |
| RNase-Free Water | ukuya kuthi ga kwi-25μL |
Amanqaku: Umthamo wokuphendula yi-10-50μL.
2. UMgaqo woMsebenzi weBhayisekile (Sumgangatho)
| Umjikelo inyathelo | Temp. | Ixesha | Imijikelo |
| Reverse Transcription | 55 ℃ | 10 imiz | 1 |
| I-Denaturation yokuqala | 95 ℃ | 30 imizuzwana | 1 |
| I-Denaturation | 95 ℃ | 10 imizuzwana | 45 |
| Ukwandiswa/Ukwandiswa | 60 ℃ | 30 imizuzwana |
| Umgaqo weBhayisekile (Khawuleza) Umjikelo inyathelo |
Temp. |
Ixesha |
Imijikelo |
| Reverse Transcription | 55 ℃ | 5 imiz | 1 |
| I-Denaturation yokuqala | 95 ℃ | 5 s | 1 |
| I-Denaturation | 95 ℃ | 3 s | 43 |
| Ukwandiswa/Ukwandiswa | 60 ℃ | 10 s |
Amanqaku:
a.Ubushushu bombhalo obubuyisela umva buphakathi kwe-50 ℃ ukuya kwi-60 ℃, ukwandisa ubushushu kunceda ukukhulisa izakhiwo ezintsonkothileyo kunye neetemplates eziphezulu zeCG;
b.Elona qondo liphezulu lobushushu bokucoca kufuneka lihlengahlengiswe ngokusekelwe kwixabiso le-Tm le-primer, kwaye ukhethe elona xesha lifutshane lokuqokelela isiginali ye-fluorescence esekelwe kwi-Real Time PCR isixhobo.
Amanqaku
Nceda unxibe iPPE eyimfuneko, idyasi yelebhu kunye neeglavu, ukuqinisekisa impilo nokhuseleko lwakho!
