(96-neza) Byihuta Byoroshye Akagari RT-qPCR Kit - Taqman
Ibisobanuro
Uwiteka(96-neza) ByihuseTM Akagari ka RT-qPCR Kit-Taqmanitangasisitemu idasanzwe ya lysisto kurekura vuba RNAkuva mu kagari k’umucoingero za reaction ya RT-qPCR, ikuraho igihe kandi ikora cyaneUburyo bwo kweza RNA, kandi bifata iminota 7 gusa kugirango ubone icyitegererezo cya RNA.Uwiteka5 × Direct RT ivanzena2 × Direct qPCR ivanze-SYBRbyatanzwe mu gasanduku birashobora kwihuta kandikubona neza igihe nyacyoIbisubizo bya PCR.
5 × Direct RT ivanze na 2 × Direct qPCR Ivanga-Taqman ifite kwihanganira imbaraga za inhibitor, kandi irashobora gukoresha lysate yicyitegererezo kugirango igeragezwe nkicyitegererezo cyo kwandukura neza no kwongerera imbaraga.Reagent irimo Foregene idasanzwe ya Reverse Transcriptase ifitanye isano cyane na RNA, hamwe na Hoteri D- Taq DNA Polymerase, DNTPs, MgCl2 , reaction reaction, PCR optimizer na stabilisateur, kandi irashobora gukoreshwa ifatanije na lysis buffer kugirango tumenye icyitegererezo vuba, byoroshye kandi neza, kandi ifite ibiranga sensibilité yo hejuru, umwihariko ukomeye kandi uhamye.
Igikoresho kigenewe micro-sisitemu lysis ya selile 96 zifite imico myiza, kandi ifite uburinganire bwiza kandi buhoraho;ibikoresho bigize ibikoresho bitanga ibisubizo 96 bya lysis, 96 reaction yo kwandukura hamwe na 96 × 2 qPCR, bishobora guhura na plaque ya selile 96 kugirango ikoreshwe rimwe, birinda umwanda uterwa no gufungura inshuro nyinshi, gukonjesha no gukonjesha reagent no gutesha agaciro imikorere ya reagent.
Ibigize ibikoresho
| Ibigize ibikoresho ( 50 μL lysis sisitemu / 20 μLRT sisitemu yo kwitwara / 20μL qPCR sisitemu yo kwitwara) | DRT-03021 | Ongera wibuke | |
| 96T | |||
| IgiceI | BufferCL | 5 mL | AkagariLysis |
| ForegeneProtease Yongeyeho II | 100 μL | ||
| BufferST | 500 μL | ||
| Igice II | ADNEraser | 100 μL | |
| 5 × Direct RT ivanze | 400 μL | RT | |
| 2 × Kuvanga qPCR-Taqman | 1 mL × 2 | QPCR | |
| 50 × ROX Yerekana irangi | 400µL | ||
| RNase- UbuntuddH2 O | 1.7mL |
| |
| Mburi mwaka | 1 igice | 1 gukorera | |
*: Lysis reagent ADN Eraser iri mubice II byigikoresho;Cell Lysis, RT, na qPCR birashobora kugurwa ukundi.
Ibiranga ibyiza
■Byoroshye kandi byiza: hamwe na tekinoroji ya Cell Direct RT, ingero za RNA zirashobora kuboneka muminota 7 gusa.
Icyitegererezo gikenewe ni gito, nkuko selile 10 zishobora kugeragezwa.
Thr Kwinjiza cyane: irashobora kumenya vuba RNA mu ngirabuzimafatizo zifite 384, 96, 24, 12, 6-amasahani meza.
Er ADN Eraser irashobora gukuraho vuba genome zasohotse, bikagabanya cyane ingaruka kubisubizo byakurikiyeho.
System Optimized RT na qPCR sisitemu ituma intambwe ebyiri RT-PCR ihinduranya transcript neza kandi PCR ikarushaho kuba nziza, kandi ikarwanya RT-qPCR inhibitor.
Porogaramu
Igipimo cyo gushyira mu bikorwa: ingirabuzimafatizo.
- RNA yarekuwe na sample lysis: ikoreshwa gusa kuri RT-qPCR inyandikorugero yiki gitabo.
- Igikoresho kirashobora gukoreshwa mubikorwa bikurikira: gusesengura imvugo ya gene, kugenzura ingaruka zo gucecekesha gene ya siRNA, gusuzuma ibiyobyabwenge, nibindi.
Ububiko hamwe nubuzima bwa Shelf
Igice cya I cyiki gitabo kigomba kubikwa kuri 4℃;Igice cya II kigomba kubikwa kuri -20 ℃.
Foregene Protease Plus II igomba kubikwa kuri 4, Ntugahagarike kuri -20 ℃.
Reagent 2×QPCR itaziguye ivanze-Taqman igomba kubikwa kuri -20℃mu mwijima;iyo ikoreshejwe kenshi, irashobora kandi kubikwa kuri 4Kubika igihe gito (koresha muminsi 10).
Igihe nyacyo PCR primer igishushanyo mbonera
Imbere Primer na Reverse Primer
Kubihe Byukuri PCR, igishushanyo mbonera ni ngombwa cyane.Ibanze bifitanye isano nuburyo bwihariye bwo kongera PCR, kandi birashobora gushushanywa hifashishijwe amahame akurikira:
- Uburebure bwa primer: 18-30bp.
- Ibirimo GC: 40-60%.
- Agaciro ka Tm: Porogaramu ishushanya, nka Primer 5, irashobora gutanga Tm agaciro ka primer.Indangagaciro za Tm zo hejuru no kumanuka primers zigomba kuba hafi bishoboka.Inzira yo kubara ya Tm irashobora kandi gukoreshwa: Tm = 4 ° C (G + C) + 2 ° C (A + T).Iyo ukora PCR, ubushyuhe buri munsi ya primer Tm agaciro ka 5 ° C muri rusange byatoranijwe nkubushyuhe bwa annealing (kwiyongera gukwiranye nubushyuhe bwa annealing birashobora kongera umwihariko wa reaction ya PCR).
- Ibicuruzwa nibicuruzwa bya PCR:
- Gushushanya primer PCR yongerera ibicuruzwa uburebure nibyiza 100-150bp.
- Igishushanyo mbonera mugice cya kabiri cyubatswe cyicyitegererezo kigomba kwirindwa bishoboka.
- Irinde gushiraho ibice 2 cyangwa byinshi byuzuzanya hagati ya 3 ′ impera yimbere na epfo na ruguru.
- Primer 3 ′ itumanaho ntirishobora kuboneka hamwe na 3 yongeyeho G cyangwa C.
- Primers ubwayo ntishobora kugira ibyuzuzanya, bitabaye ibyo imiterere yimisatsi izashyirwaho, bigira ingaruka kuri PCR.
- ATCG igomba gukwirakwizwa uko bishoboka kwose murwego rwa primer, kandi 3 ′ base base igomba kwirindwa nka T.
Umugereka1 ： C.ellRT-qPCR Kit component
1.Cell Lysis Igisubizo
| Akagari ka Lysis | |||
| Ibigize ibikoresho (24-neza lysis sisitemu / neza) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T. | 500 T. | ||
| IgiceI. | Buffer CL | 20 ml | 100 ml |
| Foregene Protease Yongeyeho II | 400 μl | 1 ml × 2 | |
| Buffer ST | 1 ml × 2 | 10 ml | |
| IgiceII | Gusiba ADN | 400 μl | 1 ml × 2 |
| RT Kuvanga | |
| Ibigize ibikoresho (20 μl sisitemu yo kwitwara) | DRT-01011-B1 |
| 200 T. | |
| 5 × Direct RT ivanze | 800 μl |
| Ubusa ddH2O | 1,7 ml × 2 |
| QPCR Kuvanga | ||
| Ibigize ibikoresho (20 μl sisitemu yo kwitwara) | DRT-01021-C1 | DRT-01021-C2 |
| 200 T. | 1000 T. | |
| 2 × Direct qPCR Ivanga-Taqman | 1 ml × 2 | 1,7 ml × 6 |
| 20 × Irangi ryerekana irangi | 40 μl | 200 μl |
| Ubusa ddH2O | 1,7 ml | 10 ml |
Imfashanyigisho:
