Enesimo sengqondo esilungileyo nesinenkqubela kumdla wabathengi, umbutho wethu uhlala uphucula umgangatho weemveliso zethu ukwanelisa iimfuno zabathengi kwaye ugxile ngakumbi kukhuseleko, ukuthembeka, ukucaciswa kokusingqongileyo, kunye nokuveliswa kweFactory Supply yaseChina eLungileyo kunye neRt-PCR eKhawulezayo Dspathtm 4X iNyathelo elinye leMultiplex Master Mix, Inkxaso yethu yonke inokuba ngamava okusebenza.Siyakwamkela ngokunyanisekileyo ukuba umise iwebhusayithi yethu kunye nenkampani kwaye usithumelele umbuzo wakho.
Enesimo sengqondo esilungileyo nesinenkqubela kumdla wabathengi, umbutho wethu uhlala uphucula umgangatho weemveliso zethu ukuze ukwanelisa iimfuno zabathengi kwaye ugxile ngakumbi kukhuseleko, ukuthembeka, ukucaciswa kokusingqongileyo, kunye nokusungula izinto ezintsha.China Taq DNA Polymerase, Qpcr, Ngoku sinezisombululo ezingcono kakhulu kunye neentengiso zezakhono kunye neqela lobugcisa.Ngophuhliso lwenkampani yethu, siyakwazi ukubonelela abathengi ngemveliso engcono kakhulu, inkxaso yobugcisa obuhle, inkonzo egqibeleleyo emva kokuthengisa.

Iinkcazelo

Le khithi isebenzisa inkqubo ye-lysis buffer ekhethekileyo enokuthi ikhulule ngokukhawuleza i-RNA kwiisampulu zeseli ezikhuliswe kwiimpendulo ze-RT-qPCR, ngaloo ndlela iphelisa inkqubo yokucoca i-RNA ethatha ixesha kunye nenzima.Ithemplethi ye-RNA inokufumaneka kwimizuzu nje esi-7.Umxube we-RT oyi-5 × ngokuthe ngqo kunye ne-2 × Direct qPCR Mix-SYBR ii-reagents ezibonelelwe yikhithi zinokufumana ngokukhawuleza nangempumelelo iziphumo zePCR zexesha lokwenyani.

I-5 × Direct RT Mix kunye ne-2 × Direct qPCR Mix-SYBR inokunyamezela inhibitor eqinile, kunye ne-lysate yeesampuli zingasetyenziswa njenge template ye-RT-qPCR ngokuthe ngqo.Le khithi iqulethe i-RNA ephezulu ehambelana ne-Foregene reverse transcriptase, kunye ne-Hot D-Taq DNA polymerase, i-dNTPs, i-MgCl₂, i-reaction buffer, i-PCR optimizer kunye ne-stabilizer.

Iinkcukacha

200×20μl Rxns, 1000×20μl Rxns

Amacandelo ekhithi

Iimpawu&izinto eziluncedo

■ Ilula kwaye isebenzayo : ngeteknoloji ye-Cell Direct RT, iisampuli ze-RNA zinokufumaneka kwimizuzu nje eyi-7.

∎ Imfuno yesampulu incinci, ingaphantsi njengeeseli ezili-10 zinokuvavanywa.

■ Ukugqithisa okuphezulu: inokubona ngokukhawuleza i-RNA kwiiseli ezikhuliswe kwii-384, 96, 24, 12, 6-well plates.

■ I-DNA Eraser inokususa ngokukhawuleza iigenomes ezikhutshiweyo, inciphise kakhulu impembelelo kwiziphumo zovavanyo ezilandelayo.

■ Inkqubo elungiselelwe i-RT kunye ne-qPCR yenza amanyathelo amabini e-RT-PCR akhutshelwe umva asebenze ngakumbi kwaye i-PCR icace ngakumbi, kwaye imelane ngakumbi ne-RT-qPCR reaction inhibitors.

Isicelo seKit

Ububanzi besicelo: iiseli ezikhuliswe.

- I-RNA ekhutshwe ngesampuli ye-lysis: isebenza kuphela kwi-template ye-RT-qPCR yale khithi.

- Ikiti ingasetyenziselwa ezi njongo zilandelayo: uhlalutyo lwe-gene expression, ukuqinisekiswa kwe-siRNA-mediated gene silence effect, ukuhlolwa kweziyobisi, njl.

Umzobo

Ugcino kunye nobomi beShelf

Inxalenye I yale khithi kufuneka igcinwe kwi-4℃;Icandelo II kufuneka ligcinwe kwi -20 ℃.

I-Foregene Protease Plus II kufuneka igcinwe ku-4℃, ungakhenkcezisi ku -20℃.

I-Reagent 2 × Direct qPCR Mix-SYBR kufuneka igcinwe kwi -20 ℃ ebumnyameni;ukuba isetyenziswe rhoqo, inokugcinwa kwi-4 ℃ yokugcina ixesha elifutshane (ukusetyenziswa ngaphakathi kweentsuku ze-10) .Enesimo sengqondo esilungileyo kunye nenkqubela phambili kumdla womthengi, umbutho wethu uphucula ngokuqhubekayo umgangatho weemveliso zethu ukwanelisa iimfuno zabathengi kwaye ugxininise ngakumbi kukhuseleko, ukuthembeka, ukucaciswa kokusingqongileyo, kunye nokutsha kweFactory Supply Chinat-Multixt-RXTMR I-Mixt-RXT i-Quick-RXT i-Mixt-RXT i-Mixt-RXT i-Quick-Right Master, i-DPP, i-DPA, i-Quick, i-Mixt-RXT, i-DPA, i-Quick, i-Mixt-Rxt, i-Mixt-RXT, i-Quick, i-DPA, i-Mixt-RXT, i-Quick, i-DPA, i-Mixt-RXT, i-Disk. abasebenzi abanamava antsokothileyo banokusuka entliziyweni kwinkxaso yakho.Siyakwamkela ngokunyanisekileyo ukuba umise iwebhusayithi yethu kunye nenkampani kwaye usithumelele umbuzo wakho.
Ubonelelo lwefektriChina Taq DNA Polymerase, Qpcr, Ngoku sinezisombululo ezingcono kakhulu kunye neentengiso zezakhono kunye neqela lobugcisa.Ngophuhliso lwenkampani yethu, siyakwazi ukubonelela abathengi ngemveliso engcono kakhulu, inkxaso yobugcisa obuhle, inkonzo egqibeleleyo emva kokuthengisa.

Ixesha lokwenyani imigaqo yoyilo lwe-PCR primer

I-Primer yokuPhambili kunye ne-Primer yokuBuyisa

Ngexesha lokwenyani iPCR, uyilo lweprimer lubaluleke kakhulu.Iiprimers zinxulumene nobucacileyo kunye nokusebenza kakuhle kokwandiswa kwePCR, kwaye zinokuyilwa ngokubhekiselele kule migaqo ilandelayo:

Ubude bePrimer: 18-30bp.

Umxholo weGC: 40-60%.

Ixabiso le-Tm: I-software yoyilo lwe-Primer, njenge-Primer 5, inokunika ixabiso le-Tm le-primer.Amaxabiso e-Tm ee-primers ezinyukayo nezisezantsi kufuneka zibe kufutshane kangangoko.Ifomula yokubala ye-Tm nayo ingasetyenziswa: Tm = 4 °C (G + C) + 2 °C (A + T).Xa kusenziwa i-PCR, iqondo lobushushu elingaphantsi kwexabiso le-primer Tm le-5 °C likhethwa ngokubanzi njengeqondo lobushushu lokutshisa (ukunyuka okuhambelanayo kwiqondo lobushushu lobushushu kunokunyusa ukuchana kwe-PCR reaction).

Iiprimers kunye neemveliso zePCR:

I-design primer PCR yokukhulisa ubude bemveliso ikhethwa yi-100-150bp.

Ii-primers zoyilo kwindawo yesibini yesakhiwo setemplate kufuneka igwenywe kangangoko kunokwenzeka.

Gwema ukubunjwa kweziseko ezi-2 okanye ngaphezulu ezihambelanayo phakathi kwee-3′ zeziphelo ze-primers ezinyukayo kunye nezantsi.

Isiseko se-Primer 3′ asikwazi ukubakho nge-3 eyongezelelweyo elandelelanayo i-G okanye i-C.

Iiprimers ngokwazo azikwazi ukuba nezakhiwo ezincedisayo, ngaphandle koko kuya kwenziwa isakhiwo se-hairpin, esichaphazela ukukhulisa i-PCR.

I-ATCG kufuneka isasazwe ngokulinganayo ngokulandelelana kwe-primer, kwaye isiseko se-terminal ye-3 kufuneka sigwenywe njenge-T.

ISihlomelo 1: Ipakethi yecandelo leCell Direct RT-qPCR Kit

1.Iseli Lysis Solution