(96-zvakanaka) QuickEasy Cell Yakananga RT-qPCR Kit - Taqman
Tsanangudzo
The(96-zvakanaka) QuickEasyTM Sero Yakananga RT-qPCR Kit-Taqmaninopayakasarudzika lysis buffer systemto nekukurumidza kuburitsa RNAkubva musero yetsikasamples dzeRT-qPCR maitiro, kubvisa iyo inopedza nguva uye yakaomaRNA yekuchenesa maitiro, uye inongotora maminetsi manomwe kuti uwane inodiwa RNA template.The5 × Yakananga RT Musanganiswauye2 × Yakananga qPCR Musanganiswa-SYBRinopiwa mubhokisi inogona kukurumidza uyezvinobudirira kuwana chaiyo-nguva uwanduPCR zvawanikwa.
5 × Yakananga RT Musanganisi uye 2 × Yakananga qPCR Mix-Taqman ine yakasimba inhibitor kushivirira, uye inogona kushandisa lysate yemuenzaniso kuti iongororwe se template yeanoshanda reverse transcription uye yakananga amplification.Iyo reagent ine Foregene yakasarudzika Reverse Transcriptase ine yakakwirira hukama hweRNA, pamwe neHot D-Taq DNA Polymerase, dNTPs, MgCl.2 , reaction buffer, PCR optimizer uye stabilizer, uye inogona kushandiswa pamwe chete ne lysis buffer kuti ione sampuli nokukurumidza, nyore uye yakarurama, uye ine maitiro ekunzwa kwakanyanya, kusimba kwakasimba uye kugadzikana kwakanaka.
Iyo kit inotarisirwa kune micro-system lysis ye96-yakanaka masero akagadzirwa, uye ane yakanaka kufanana uye kuenderana;zvikamu zvekiti zvinopa 96 lysis reactions, 96 reverse transcription reactions uye 96 × 2 qPCR reactions, iyo inogona kusangana ne96-tsime masero ndiro yekushandiswa kwekamwe chete, kudzivisa kusvibiswa kunokonzerwa nekudzokorora kuvhurika, kutonhora uye kunyunguduka kwemareagents uye kuora kwekuita reagent.
Kit zvikamu
| Kit composition ( 50 μL lysis system/20 μLRT maitiro system /20μL qPCR reaction system) | DRT-03021 | Remark | |
| 96T | |||
| ChikamuI | BufferCL | 5 mL | SeroLysis |
| ForegeneProtease Plus II | 100 μL | ||
| BufferST | 500 μL | ||
| Chikamu II | DNAEraser | 100 μL | |
| 5 × Yakananga RT Musanganiswa | 400 μL | RT | |
| 2 × Yakananga qPCR Musanganiswa-Taqman | 1 mL × 2 | qPCR | |
| 50 × ROX Reference Dye | 400µL | ||
| RNase- FreeddH2 O | 1.7mL |
| |
| Manual | 1 chidimbu | 1 kushumira | |
*: Iyo lysis reagent DNA Eraser inosanganisirwa muChikamu II chekiti;Cell Lysis, RT, uye qPCR zvikamu zvinogona kutengwa zvakasiyana.
Zvimiro&zvakanakira
■Yakareruka uye inoshanda: neCell Direct RT tekinoroji, RNA samples inogona kuwanikwa mumaminetsi manomwe chete.
∎ Sample kudiwa idiki, yakaderera semaseru gumi anogona kuongororwa.
■ High throughput: inogona kukurumidza kuona RNA mumaseru akagadzirwa mu384, 96, 24, 12, 6-tsime mahwendefa.
∎ DNA Eraser inogona kukurumidza kubvisa majenomes akaburitswa, kuderedza zvakanyanya kukanganiswa pamhedzisiro yekuedza.
■ Optimized RT uye qPCR sisitimu inoita kuti nhanho mbiri RT-PCR reverse transcription inyatsoita uye PCR yakanyatsojeka, uye inoshingirira kuRT-qPCR reaction inhibitors.
Kit application
Kuwanda kwekushandiswa: masero akagadzirwa.
- RNA yakaburitswa nesample lysis: inoshanda chete kune RT-qPCR template yeiyi kit.
- Iyo kit inogona kushandiswa kune zvinotevera zvinangwa: gene kutaura kuongororwa, kuoneswa kweSiRNA-mediated gene kunyaradza maitiro, kuongorora zvinodhaka, nezvimwe.
Storage uye Sherufu upenyu
Chikamu I chekiti ichi chinofanira kuchengetwa pa4℃;Chikamu II chinofanira kuchengetwa pa -20 ℃.
Foregene Protease Plus II inofanira kuchengetwa pa4℃, usaome pa -20 ℃.
Reagent 2×Yakananga qPCR Mix-Taqman inofanira kuchengetwa pa -20℃murima;kana ikashandiswa kazhinji, inogona zvakare kuchengetwa pa4℃ yekuchengetedza kwenguva pfupi (shandisa mukati memazuva gumi).
Real Time PCR primer dhizaini misimboti
Pamberi Primer uye Reverse Primer
Kune Real Time PCR, dhizaini yekutanga yakakosha.MaPrimers ane hukama nehunyanzvi uye kugona kwePCR amplification, uye inogona kugadzirwa ichitarisa kune anotevera misimboti:
- Kureba kwekutanga: 18-30bp.
- GC yemukati: 40-60%.
- Tm kukosha: Primer dhizaini software, senge Primer 5, inogona kupa iyo Tm kukosha kweiyo primer.Iyo Tm tsika dzekumusoro uye pasi pemvura dzekutanga dzinofanira kunge dziri padyo sezvinobvira.The Tm calculation formula inogonawo kushandiswa: Tm = 4 °C (G + C) + 2 °C (A + T).Paunenge uchiita PCR, tembiricha iri pasi peiyo primer Tm kukosha kwe5 ° C inowanzo sarudzwa seye tembiricha yekudziya (iyo inowirirana kuwedzera kwekushisa kweanona inogona kuwedzera iyo chaiyo yePCR reaction).
- Primers uye PCR zvigadzirwa:
- Dhizaini primer PCR yekusimudzira chigadzirwa kureba ingangoita 100-150bp.
- Dhizaini yekutanga munzvimbo yechipiri yemaitiro eiyo template inofanira kudziviswa zvakanyanya sezvinobvira.
- Dzivisa kuumbwa kwe 2 kana anopfuura mabhesi anowirirana pakati pe 3′ migumo yekumusoro nekudzika kwemvura.
- Primer 3′ terminal base haigone kuvepo ne3 yekuwedzera yakatevedzana G kana C.
- Iwo maprimer pachawo haagone kuve neanopindirana zvimiro, zvikasadaro chimiro chebvudzi chinoumbwa, chinokanganisa PCR amplification.
- ATCG inofanirwa kugovaniswa zvakaenzana sezvinobvira mukutevedzana kwekutanga, uye 3 ′ terminal base inofanira kudzivirirwa seT.
Appendikisi1:Cuye DirectRT-qPCR Kit component supplement pack
1.Cell Lysis Solution
| Sero Lysis Solution | |||
| Kit zvikamu (24-tsime lysis system / tsime) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T | 500 T | ||
| ChikamuI | Buffer CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 μl | 1 ml × 2 | |
| Buffer ST | 1 ml × 2 | 10 ml | |
| ChikamuII | DNA Eraser | 400 μl | 1 ml × 2 |
| RT Mix | |
| Kit zvikamu (20 μl reaction system) | DRT-01011-B1 |
| 200 T | |
| 5 × Yakananga RT Musanganiswa | 800 μl |
| RNase-Yemahara ddH2O | 1.7 ml × 2 |
| qPCR Mix | ||
| Kit zvikamu (20 μl reaction system) | DRT-01021-C1 | DRT-01021-C2 |
| 200 T | 1000 T | |
| 2 × Yakananga qPCR Mix-Taqman | 1 ml × 2 | 1.7 ml × 6 |
| 20 × ROX Reference Dye | 40 μl | 200 μl |
| RNase-Yemahara ddH2O | 1.7 ml | 10 ml |
Mirayiridzo yezvinyorwa:
