Ibyo bifite imyumvire myiza kandi itera imbere kubwinyungu zabakiriya, umuryango wacu uhora utezimbere ubuziranenge bwibicuruzwa byacu kugirango uhaze ibyifuzo byabaguzi kandi ukomeza kwibanda ku mutekano, kwiringirwa, ibisobanuro by’ibidukikije, no guhanga udushya two gutanga ibicuruzwa mu Bushinwa Byoroshye kandi byihuse Rt-PCR Dspathtm 4X Intambwe imwe ya Multiplex Master Mix, Abakozi bacu bafite uburambe burashobora kuba babikuye ku mutima ku nkunga yawe.Turabashimira byimazeyo guhagarika byanze bikunze kurubuga rwacu hamwe nisosiyete no kutwoherereza ibibazo byanyu.
Ibyo bifite imyumvire myiza kandi itera imbere kubwinyungu zabakiriya, umuryango wacu uhora utezimbere ibicuruzwa byacu kugirango uhaze ibyifuzo byabaguzi kandi ukomeza kwibanda kumutekano, kwiringirwa, ibidukikije, no guhanga udushya.Ubushinwa Taq ADN Polymerase, Qpcr, Ubu dufite ibisubizo byiza hamwe no kugurisha ubuhanga hamwe nitsinda rya tekiniki. Hamwe niterambere ryikigo cyacu, turashobora guha abakiriya ibicuruzwa byiza, inkunga nziza ya tekiniki, serivisi nziza nyuma yo kugurisha.

Ibisobanuro

Iki gikoresho gikoresha sisitemu idasanzwe ya lysis buffer ishobora kurekura byihuse RNA mumikorere yimikorere yimikorere ya RT-qPCR, bityo bikuraho inzira yo kweza RNA itwara igihe kandi ikora cyane.Inyandikorugero ya RNA irashobora kuboneka muminota 7 gusa.5 × Direct RT ivanze na 2 × Imiyoboro ya qPCR ivanze-SYBR itangwa nigitabo irashobora kwihuta kandi neza kubona ibisubizo nyabyo-byuzuye bya PCR.

5 × Direct RT ivanze na 2 × Direct qPCR ivanze-SYBR ifite kwihanganira imbaraga za inhibitor, kandi lysate yintangarugero irashobora gukoreshwa nkicyitegererezo cya RT-qPCR itaziguye.Iki gikoresho kirimo RNA idasanzwe-yuzuye ya Foregene reverse transcriptase, hamwe na polymerase ya D-Taq ishyushye, DNTPs, MgCl₂, reaction ya reaction, optimizer ya PCR na stabilisateur.

Ibisobanuro

200 × 20μl Rxns, 1000 × 20μl Rxns

Ibigize ibikoresho

Ibiranga ibyiza

■ Byoroshye kandi byiza: hamwe na tekinoroji ya Direct 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.

Igishushanyo

Ububiko hamwe nubuzima bwa Shelf

Igice cya I cy'iki gikoresho kigomba kubikwa kuri 4 ℃;Igice cya II kigomba kubikwa kuri -20 ℃.

Foregene Protease Plus II igomba kubikwa kuri 4 ℃, ntugahagarike kuri -20 ℃.

Reagent 2 × Direct qPCR Ivanga-SYBR igomba kubikwa kuri -20 ℃ mu mwijima;iyo ikoreshejwe kenshi, irashobora kandi kubikwa kuri 4 ℃ kubikwa mugihe gito (koresha mugihe cyiminsi 10) .Iyo ifite imyumvire myiza kandi itera imbere kubyifuzo byabakiriya, umuryango wacu uhora utezimbere ibicuruzwa byacu kugirango uhaze ibyifuzo byabaguzi kandi ukomeza kwibanda kumutekano, kwiringirwa, kubidukikije, no guhanga udushya twibikorwa byinganda Byoroshye kandi byoroshye Rt-PCR Dspathtm 4X One-Intambwe Yuzuye.Turabashimira byimazeyo guhagarika byanze bikunze kurubuga rwacu hamwe nisosiyete no kutwoherereza ibibazo byanyu.
Gutanga UrugandaUbushinwa Taq ADN Polymerase, Qpcr, Ubu dufite ibisubizo byiza hamwe no kugurisha ubuhanga hamwe nitsinda rya tekiniki. Hamwe niterambere ryikigo cyacu, turashobora guha abakiriya ibicuruzwa byiza, inkunga nziza ya tekiniki, serivisi nziza nyuma yo kugurisha.

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.

Umugereka 1 ： Akagari kayobora RT-qPCR Ibikoresho byuzuza ibikoresho

1.Akagari ka Lysis