Cell Direct RT qPCR Kit-Taqman Direct Cell Lysis Cell Yakagadzirira Imwe-nhanho qRT-PCR Kits Probe
Tsanangudzo
Ichi chigadzirwa chinoshandisa yakasarudzika lysis buffer sisitimu kukurumidza kuburitsa RNA kubva kune yakasimwa maseru masampula eRT-qPCR maitiro, kubvisa iyo inopedza nguva uye inorema RNA yekuchenesa maitiro, uye chete maminetsi manomwe kuwana inodiwa RNA template, ine 5 × Direct RT Musanganiswa, 2 × Direct qPCR Mix-Taqman inopiwa neiyo kit inogoneka-inoshanda PC.
5 × Yakananga RT Musanganisi uye 2 × Yakananga qPCR Mix-Taqman ine yakasimba inhibitor kushivirira uye inogona kuita zvinobudirira kudzosera uye yakananga amplification uchishandisa lysate yesampuli kuyerwa setemplate.Iyo reagent ine Foregene Reverse Transcriptase, Inopisa D-Taq DNA Polymerase, dNTPs, MgCl.2, Reaction Buffer, PCR Optimizer uye Stabilizer, iyo inogona kushandiswa ne lysis buffer kukurumidza uye nyore kuona samples, uye ine hunhu hwekunzwa kwakanyanya, hutsanana uye kugadzikana.
Zvinotsanangurwa
200×20μl Rxns, 1000×20μl Rxns
Kit zvikamu
| QuickEasyTMSero Yakananga RT-qPCR Kit-Taqman | ||||
| Kit zvikamu 20μl qPCR Reaction System | DRT-01021 | DRT-01022 | Cherechedza | |
| 200 T | 1000 T | |||
|
Chikamu I | Buffer CL | 4 ml | 20 ml | Sero Lysis |
| Foregene Protease Plus II | 80 μl | 400 μl | ||
| Buffer ST | 400 μl | 1 ml × 2 | ||
|
Chikamu II | DNA Eraser | 80 μl | 400 μl | |
| 5×Yakananga RT Musanganiswa * | 160 μl | 800 μl | RT | |
| 2 × Yakananga qPCR Musanganiswa-Taqman * | 1 ml × 2 | 1.7 ml × 6 | qPCR | |
| 20 × ROX Reference Dye | 40 μl | 200 μl | ||
| RNase-Yemahara ddH2O | 1.7 ml | 10 ml | ||
| Bhuku reMirayiridzo | 1 chidimbu | 1 chidimbu | ||
*:Cell Lysis, 5 × Direct RT Musanganiswa, 2 × Yakananga qPCR Mix-Taqman inogona 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:
