Umbutho ugcina ingcamango yenkqubo "ulawulo lwesayensi, umgangatho ophezulu kunye nokusebenza kakuhle, umthengi ophakamileyo we-China Manufacturer ye-2 × i-Concentrated Premix yeSampuli engahlanjululwanga ye-Real-Time Quantitative PCR Direct Multiplex Probe Qpcr Mix Plus U, Ishishini lethu lizinikele ekunikeni abathengi ngeyona nto ibalulekileyo kwaye ikhuselekile iimveliso eziphezulu zomthengi ngexabiso ngalinye elinobunkunkqele kunye nexabiso eliphakamileyo.
Umbutho ugcina ingqikelelo yenkqubo "ulawulo lwesayensi, umgangatho ophezulu kunye nokusebenza kakuhle, umthengi ophakamileyoI-China Taq DNA Polymerase kunye ne-Qpcr, Inkampani yethu inamandla amaninzi kwaye inenkqubo yokuthengisa ezinzileyo negqibeleleyo.Sinqwenela ukuba siseke ubudlelwane beshishini obuvakalayo nabo bonke abathengi abasuka ekhaya nakumazwe aphesheya ngokwesiseko seenzuzo ezifanayo.
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:

1. I-design primer PCR yokukhulisa ubude bemveliso ikhethwa yi-100-150bp.
2. Ii-primers zoyilo kwindawo yesibini yesakhiwo setemplate kufuneka igwenywe kangangoko kunokwenzeka.
3. Gwema ukubunjwa kweziseko ezi-2 okanye ngaphezulu ezihambelanayo phakathi kwee-3′ zeziphelo ze-primers ezinyukayo kunye nezantsi.
4. Isiseko se-Primer 3′ asikwazi ukubakho nge-3 eyongezelelweyo elandelelanayo i-G okanye i-C.
5. Iiprimers ngokwazo azikwazi ukuba nezakhiwo ezincedisayo, ngaphandle koko kuya kwenziwa isakhiwo se-hairpin, esichaphazela ukukhulisa i-PCR.
6. I-ATCG kufuneka isasazwe ngokulinganayo ngokulandelelana kwe-primer, kwaye isiseko se-terminal ye-3 kufuneka sigwenywe njenge-T.

Isihlomelo1:Cell ngqoI-RT-qPCR Ikhithi icandeloIpakethi yokongeza

1.Cell Lysis Solution

 Umbutho ugcina ingcamango yenkqubo "ulawulo lwesayensi, umgangatho ophezulu kunye nokusebenza kakuhle, umthengi ophakamileyo we-China Manufacturer ye-2 × i-Concentrated Premix yeSampuli engahlanjululwanga ye-Real-Time Quantitative PCR Direct Multiplex Probe Qpcr Mix Plus U, Ishishini lethu lizinikele ekunikeni abathengi ngeyona nto ibalulekileyo kwaye ikhuselekile iimveliso eziphezulu zomthengi ngexabiso ngalinye elinobunkunkqele kunye nexabiso eliphakamileyo.
China Manufacturer forI-China Taq DNA Polymerase kunye ne-Qpcr, Inkampani yethu inamandla amaninzi kwaye inenkqubo yokuthengisa ezinzileyo negqibeleleyo.Sinqwenela ukuba siseke ubudlelwane beshishini obuvakalayo nabo bonke abathengi abasuka ekhaya nakumazwe aphesheya ngokwesiseko seenzuzo ezifanayo.

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:

1. I-design primer PCR yokukhulisa ubude bemveliso ikhethwa yi-100-150bp.
2. Ii-primers zoyilo kwindawo yesibini yesakhiwo setemplate kufuneka igwenywe kangangoko kunokwenzeka.
3. Gwema ukubunjwa kweziseko ezi-2 okanye ngaphezulu ezihambelanayo phakathi kwee-3′ zeziphelo ze-primers ezinyukayo kunye nezantsi.
4. Isiseko se-Primer 3′ asikwazi ukubakho nge-3 eyongezelelweyo elandelelanayo i-G okanye i-C.
5. Iiprimers ngokwazo azikwazi ukuba nezakhiwo ezincedisayo, ngaphandle koko kuya kwenziwa isakhiwo se-hairpin, esichaphazela ukukhulisa i-PCR.
6. I-ATCG kufuneka isasazwe ngokulinganayo ngokulandelelana kwe-primer, kwaye isiseko se-terminal ye-3 kufuneka sigwenywe njenge-T.

Isihlomelo1:Cell ngqoI-RT-qPCR Ikhithi icandeloIpakethi yokongeza

1.Cell Lysis Solution

Izikhokelo zoMyalelo:

 Iseli eKhawulezayo eNgqo ngqo i-RT-qPCR Kit-Taqman