(96-da kyau) Kit ɗin Mai Sauƙi Mai Sauƙi Kai tsaye RT-qPCR - SYBR Green I
Bayani
The(96-da kyau) QuickEasyTM Wayar salula Direct RT-qPCR Kit-SYBR Green Iyana bayarwana musamman lysis buffer tsarinto da sauri saki RNAdaga celled al'adasamfurori don halayen RT-qPCR, kawar da cin lokaci da aikiTsarin tsarkakewa na RNA, kuma yana ɗaukar mintuna 7 kawai don samun samfurin RNA da ake buƙata.The5× Direct RT Mixkuma2× Direct qPCR Mix-SYBRbayar a cikin akwatin iya sauri da kumayadda ya kamata samun ainihin-lokaci adadiSakamakon PCR.
5 × Direct RT Mix da 2 × Direct qPCR Mix-SYBR suna da ƙarfin juriya mai hanawa, kuma suna iya amfani da lysate na samfurin don gwadawa azaman samfuri don ingantaccen juzu'i da ƙayyadaddun haɓakawa.Reagent ya ƙunshi Foregene na musamman Reverse Transcriptase tare da babban kusanci ga RNA, da Hot D-Taq DNA Polymerase, dNTPs, MgCl.2 , Matsakaicin amsawa, PCR optimizer da stabilizer , kuma za'a iya amfani dashi tare da lysis buffer don gano samfurin da sauri, sauƙi da kuma daidai, kuma yana da halaye na babban hankali, ƙayyadaddun ƙayyadaddun ƙarfi da kwanciyar hankali mai kyau.
Kit ɗin yana nufin lysis micro-system 96-da kyau al'ada Kwayoyin, kuma yana da kyau daidaito da daidaito;Abubuwan da ke cikin kit ɗin suna ba da halayen lysis na 96, halayen juzu'i na 96 da halayen 96 × 2 qPCR, waɗanda zasu iya saduwa da farantin rijiyoyin 96-riji don amfani guda ɗaya, guje wa gurɓataccen gurɓataccen abu wanda ya haifar da maimaita buɗewa, daskarewa da narkewar reagents da lalata aikin reagent.
Abubuwan Kit
| Kayan abun ciki ( 50 μL lysis tsarin / 20 μl kuRT tsarin amsawa /20μLqPCR tsarin amsawa) | DRT-03011 | Magana | |
| 96T | |||
| SasheI | BufferCL | ml 5 | CellLysis |
| ForegeneProtease Plus II | 100 μL | ||
| BufferST | 500 μL | ||
| Sashe II | DNAGoge | 100 μL | |
| 5× Direct RT Mix | 400 μL | RT | |
| 2× Direct qPCR Mix-Farashin SYBR | 1 mL × 2 | qPCR | |
| 50× ROX Reference Dye | 400µL | ||
| RNase- KyautaddH2 O | 1.7ml |
| |
| Mshekara-shekara | 1 yanki | 1 hidima | |
*: An haɗa lysis reagent DNA Eraser a cikin Sashe na II na kit;Ana iya siyan abubuwan haɗin Cell Lysis, RT, da qPCR daban.
Fasaloli & fa'idodi
■Mai sauƙi da inganci: tare da fasahar Cell Direct RT, ana iya samun samfuran RNA a cikin mintuna 7 kawai.
∎ Bukatar samfurin karami ne, kamar yadda za a iya gwada ƙananan sel guda 10.
∎ Babban abin da ake fitarwa: zai iya gano RNA da sauri a cikin sel waɗanda aka tsara a cikin faranti 384, 96, 24, 12, 6-rijiya.
∎ Gogewar DNA na iya saurin cire kwayoyin halittar da aka fitar, da rage tasirin sakamakon gwaji na gaba.
∎ Ingantaccen tsarin RT da qPCR yana sa jujjuyawar matakai biyu na RT-PCR mafi inganci kuma PCR ta fi ƙayyadaddun ƙayyadaddun bayanai, kuma mafi juriya ga masu hana amsawar RT-qPCR.
Kit aikace-aikace
Iyakar aikace-aikace: sel masu al'ada.
- RNA da aka saki ta samfurin lysis: kawai ana amfani da samfurin RT-qPCR na wannan kit.
- Za a iya amfani da kit ɗin don dalilai masu zuwa: nazarin maganganun kwayoyin halitta, tabbatar da tasirin sirna mai tsaka-tsaki na yin shiru, gwajin magunguna, da sauransu.
Adana da Rayuwar Shelf
Ya kamata a adana sashe na I na wannan kit ɗin a 4℃;Ya kamata a adana Sashe na II a -20 ℃.
Ya kamata a adana Foregene Protease Plus II a 4℃, kar a daskare a -20 ℃.
Reagent 2×Ya kamata a adana qPCR Mix-Taqman kai tsaye a -20℃a cikin duhu;idan ana amfani dashi akai-akai, ana iya adana shi a 4℃ don ajiya na ɗan gajeren lokaci (amfani da sama a cikin kwanaki 10).
Ka'idodin ƙira na ainihin lokacin PCR
Gaba Farawa da Juya Farko
Don Real Time PCR, ƙirar farko tana da mahimmanci.Abubuwan haɓaka suna da alaƙa da ƙayyadaddun ƙayyadaddun ƙayyadaddun ƙimar PCR, kuma ana iya tsara su tare da la'akari da ƙa'idodi masu zuwa:
- Tsayin farko: 18-30bp.
- GC abun ciki: 40-60%.
- Ƙimar Tm: Software na ƙira na farko, kamar Primer 5, na iya ba da ƙimar Tm na firamare.Ma'auni na Tm na sama da na ƙasa ya kamata su kasance kusa da iyawa.Hakanan ana iya amfani da dabarar lissafin Tm: Tm = 4 °C (G + C) + 2 °C (A + T).Lokacin yin PCR, ana zaɓar zafin da ke ƙasa da ƙimar ƙimar Tm na 5 ° C gabaɗaya azaman zazzabi mai ɗaukar nauyi (madaidaicin haɓakar zafin jiki na iya ƙara ƙayyadaddun halayen PCR).
- Alamar farko da samfuran PCR:
- Tsawon samfurin PCR na ƙirar ƙira ya fi dacewa 100-150bp.
- Ya kamata a kauce wa ƙera ƙirar ƙira a cikin yanki na biyu na samfuri gwargwadon yiwuwa.
- Guji samuwar tushe guda 2 ko sama da haka tsakanin 3′ na ƙofofin sama da na ƙasa.
- Babban tushe 3′ na tashar ba zai iya kasancewa tare da ƙarin ƙarin 3 a jere G ko C.
- Masu farawa da kansu ba za su iya samun ƙarin tsari ba, in ba haka ba za a kafa tsarin gashin gashi, yana shafar haɓaka PCR.
- Yakamata a rarraba ATCG daidai gwargwado kamar yadda zai yiwu a cikin jeri na farko, kuma ya kamata a guje wa tushe na 3′ a matsayin T.
Karin bayani1:Cda DirectRT-qPCR Kit component kari kunshin
1.Cell Lysis Magani
| Cell Lysis Magani | |||
| Abubuwan Kit (24-riji tsarin lysis / rijiyar) | Saukewa: DRT-01011-A1 | Saukewa: DRT-01011-A2 | |
| 100 T | 500 T | ||
| SasheI | Buffer CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 ml | 1 ml × 2 | |
| Buffer ST | 1 ml × 2 | 10 ml | |
| SasheII | Goge DNA | 400 ml | 1 ml × 2 |
| RT Mix | |
| Abubuwan Kit (20 μl tsarin amsawa) | Saukewa: DRT-01011-B1 |
| 200 T | |
| 5× Direct RT Mix | 800 ml |
| RNase-Free ddH2O | 1.7 ml × 2 |
| qPCR Mix | ||
| Abubuwan Kit (20 μl tsarin amsawa) | Saukewa: DRT-01021-C1 | Saukewa: DRT-01021-C2 |
| 200 T | 1000 T | |
| 2× Direct qPCR Mix-Taqman | 1 ml × 2 | 1.7 ml × 6 |
| 20× ROX Reference Dye | 40ml ku | 200 μl |
| RNase-Free ddH2O | 1.7 ml | 10 ml |
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