Izi zili ndi malingaliro abwino komanso opita patsogolo ku chidwi cha kasitomala, gulu lathu limawongolera zinthu zathu mosadukiza kuti zikwaniritse zofuna za ogula ndikumayang'ananso chitetezo, kudalirika, mawonekedwe a chilengedwe, komanso luso la Factory Supply China Convenient and Quick Rt-PCR Dspathtm 4X One-Step Multiplex Master Mix, Othandizira athu atha kukhala odziwa ntchito yanu.Tikulandirani moona mtima kuti muyime pa webusayiti yathu ndi kampani yathu ndikutumiza zomwe mukufuna.
Zomwe zili ndi malingaliro abwino komanso opita patsogolo pazokonda zamakasitomala, bungwe lathu limasintha mosasintha zinthu zathu kuti zikwaniritse zofuna za ogula ndikumayang'ananso kwambiri zachitetezo, kudalirika, mawonekedwe achilengedwe, komanso ukadaulo waChina Taq DNA Polymerase, Qpcr, Tsopano tili ndi mayankho abwino kwambiri komanso malonda aluso ndi gulu laukadaulo.Pa chitukuko cha kampani yathu, timatha kupereka makasitomala abwino kwambiri, chithandizo chabwino chaukadaulo, ntchito yabwino yotsatsa pambuyo pogulitsa.

Kufotokozera

Chidachi chimagwiritsa ntchito njira yapadera ya lysis buffer yomwe imatha kumasula mwachangu RNA kuchokera ku ma cell otukuka pamachitidwe a RT-qPCR, potero amachotsa njira yoyeretsera RNA yomwe imatenga nthawi komanso yovuta.The RNA template ikhoza kupezeka mu mphindi 7 zokha.5 × Direct RT Mix ndi 2 × Direct qPCR Mix-SYBR reagents zoperekedwa ndi zida zitha kupeza mwachangu komanso moyenera zotsatira za PCR zenizeni zenizeni.

5 × Direct RT Mix ndi 2 × Direct qPCR Mix-SYBR ali ndi kulekerera kwamphamvu kwa inhibitor, ndipo ma lysate a zitsanzo angagwiritsidwe ntchito ngati template ya RT-qPCR mwachindunji.Chidachi chili ndi RNA yapamwamba kwambiri ya Foregene reverse transcriptase, ndi Hot D-Taq DNA polymerase, dNTPs, MgCl.₂, zotchingira zochita, PCR optimizer ndi stabilizer.

Zofotokozera

200×20μl Rxns, 1000×20μl Rxns

Zida zamagulu

Mbali & ubwino

■ Zosavuta komanso zothandiza : ndiukadaulo wa Cell Direct RT, zitsanzo za RNA zitha kupezeka m'mphindi 7 zokha.

■ Zofuna zachitsanzo ndizochepa, mpaka ma cell 10 akhoza kuyesedwa.

■ Kutulutsa kwakukulu: kumatha kuzindikira mwachangu RNA m'maselo opangidwa mu 384, 96, 24, 12, 6-chitsime mbale.

■ DNA chofufutira akhoza kuchotsa mwamsanga matupi athu omasulidwa, kwambiri kuchepetsa zotsatira zoyeserera.

■ Makina okhathamiritsa a RT ndi qPCR amapangitsa kuti masitepe awiri a RT-PCR alembedwe bwino kwambiri komanso PCR yachindunji, komanso yosagwirizana ndi RT-qPCR reaction inhibitors.

Kit application

Kuchuluka kwa ntchito: maselo otukuka.

- RNA yotulutsidwa ndi zitsanzo za lysis: imagwira ntchito pa template ya RT-qPCR ya zida izi.

- Zidazi zitha kugwiritsidwa ntchito pazifukwa izi: kusanthula kwa jini, kutsimikizira kwa siRNA-mediated gene silence effect, kuyang'anira mankhwala, etc.

Chithunzi

Kusungirako ndi moyo wa alumali

Gawo I la zida izi ziyenera kusungidwa pa 4 ℃;Gawo II liyenera kusungidwa pa -20 ℃.

Foregene Protease Plus II iyenera kusungidwa pa 4 ℃, osazizira pa -20 ℃.

Reagent 2 × Direct qPCR Mix-SYBR iyenera kusungidwa pa -20 ℃ mumdima;ngati agwiritsidwa ntchito pafupipafupi, amathanso kusungidwa pa 4 ℃ kuti asungidwe kwakanthawi kochepa (kugwiritsa ntchito mkati mwa masiku 10) .Zomwe zimakhala ndi malingaliro abwino komanso opita patsogolo ku chidwi cha kasitomala, gulu lathu limakweza zinthu zathu mosadukiza kuti zikwaniritse zofuna za ogula komanso limayang'ananso kwambiri zachitetezo, kudalirika, kutsimikizika kwachilengedwe, komanso luso la Factory Supply Chinat-RXTPCMRW Multipple China Convenint-RXSt. ogwira nawo ntchito odziwa zambiri akhoza kukhala ndi mtima wonse pa chithandizo chanu.Tikulandirani moona mtima kuti muyime pa webusayiti yathu ndi kampani yathu ndikutumiza zomwe mukufuna.
Factory SupplyChina Taq DNA Polymerase, Qpcr, Tsopano tili ndi mayankho abwino kwambiri komanso malonda aluso ndi gulu laukadaulo.Pa chitukuko cha kampani yathu, timatha kupereka makasitomala abwino kwambiri, chithandizo chabwino chaukadaulo, ntchito yabwino yotsatsa pambuyo pogulitsa.

Real Time PCR primer design mfundo

Forward Primer ndi Reverse Primer

Kwa Real Time PCR, mapangidwe oyambira ndi ofunika kwambiri.Zoyambira zimayenderana ndi kutsimikizika komanso kuthekera kwa kukulitsa kwa PCR, ndipo zitha kupangidwa motengera mfundo izi:

Kutalika koyambirira: 18-30bp.

Zolemba za GC: 40-60%.

Mtengo wa Tm: Mapulogalamu opangira ma primer, monga Primer 5, amatha kupereka mtengo wa Tm woyambira.Makhalidwe a Tm oyambira kumtunda ndi kumtunda ayenera kukhala pafupi momwe angathere.Njira yowerengera ya Tm ingagwiritsidwenso ntchito: Tm = 4 °C (G + C) + 2 °C (A + T).Pochita PCR, kutentha pansi pa mtengo woyambira wa Tm wa 5 ° C nthawi zambiri kumasankhidwa ngati kutentha kwapang'onopang'ono (kuwonjezeka kofananirako kwa kutentha kwapang'onopang'ono kumatha kukulitsa momwe PCR imachitira).

Primers ndi zinthu za PCR:

Design primer PCR amplification product kutalika makamaka 100-150bp.

Zoyambira zojambula m'dera lachiwiri la template ziyenera kupewedwa momwe zingathere.

Pewani kupanga zoyambira ziwiri kapena kupitilira apo pakati pa 3′ malekezero oyambira kumtunda ndi kumunsi kwa mtsinje.

Primer 3′ terminal base sangakhalepo ndi 3 zowonjezera zotsatizana za G kapena C.

Zoyambira zokha sizingakhale ndi zida zowonjezera, apo ayi mawonekedwe atsitsi adzapangidwa, zomwe zimakhudza kukulitsa kwa PCR.

ATCG iyenera kugawidwa mofanana momwe mungathere poyambira, ndipo 3′ terminal base iyenera kupewedwa ngati T.

Zowonjezera 1: Cell Direct RT-qPCR Kit chigawo chowonjezera paketi

1.Cell Lysis Solution