Ishyirahamwe rikomeza inzira yuburyo "imicungire yubumenyi, ubuziranenge bwo hejuru nubushobozi bwibanze, abaguzi basumba uwabashinwa kubucuruzi bwa 2 × Concentrated Premix for Unpurified Sample Real-Time Quantitative PCR Direct Multiplex Probe Qpcr Mix Plus U, Ubucuruzi bwacu bwihaye guha abakiriya ibicuruzwa byiza kandi bifite umutekano muke kubiciro bikaze, bigatuma buri mukiriya yishimira serivisi zacu.
Ishyirahamwe rikomeza inzira yuburyo "imiyoborere yubumenyi, ubuziranenge bwo hejuru kandi bukora neza, abaguzi bakomeye kuriUbushinwa Taq ADN Polymerase na Qpcr, Isosiyete yacu ifite imbaraga nyinshi kandi ifite sisitemu yo kugurisha ihamye kandi itunganye.Twifuzaga ko twashyiraho umubano mwiza wubucuruzi nabakiriya bose kuva mugihugu ndetse no mumahanga dushingiye kubwinyungu rusange.
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:

1. Gushushanya primer PCR yongerera ibicuruzwa uburebure nibyiza 100-150bp.
2. Igishushanyo mbonera mugice cya kabiri cyubatswe cyicyitegererezo kigomba kwirindwa bishoboka.
3. Irinde gushiraho ibice 2 cyangwa byinshi byuzuzanya hagati ya 3 ′ impera yimbere na epfo na ruguru.
4. Primer 3 ′ itumanaho ntirishobora kuboneka hamwe na 3 yongeyeho G cyangwa C.
5. Primers ubwayo ntishobora kugira ibyuzuzanya, bitabaye ibyo imiterere yimisatsi izashyirwaho, bigira ingaruka kuri PCR.
6. 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

 Ishyirahamwe rikomeza inzira yuburyo "imicungire yubumenyi, ubuziranenge bwo hejuru nubushobozi bwibanze, abaguzi basumba uwabashinwa kubucuruzi bwa 2 × Concentrated Premix for Unpurified Sample Real-Time Quantitative PCR Direct Multiplex Probe Qpcr Mix Plus U, Ubucuruzi bwacu bwihaye guha abakiriya ibicuruzwa byiza kandi bifite umutekano muke kubiciro bikaze, bigatuma buri mukiriya yishimira serivisi zacu.
Ubushinwa bukoraUbushinwa Taq ADN Polymerase na Qpcr, Isosiyete yacu ifite imbaraga nyinshi kandi ifite sisitemu yo kugurisha ihamye kandi itunganye.Twifuzaga ko twashyiraho umubano mwiza wubucuruzi nabakiriya bose kuva mugihugu ndetse no mumahanga dushingiye kubwinyungu rusange.

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:

1. Gushushanya primer PCR yongerera ibicuruzwa uburebure nibyiza 100-150bp.
2. Igishushanyo mbonera mugice cya kabiri cyubatswe cyicyitegererezo kigomba kwirindwa bishoboka.
3. Irinde gushiraho ibice 2 cyangwa byinshi byuzuzanya hagati ya 3 ′ impera yimbere na epfo na ruguru.
4. Primer 3 ′ itumanaho ntirishobora kuboneka hamwe na 3 yongeyeho G cyangwa C.
5. Primers ubwayo ntishobora kugira ibyuzuzanya, bitabaye ibyo imiterere yimisatsi izashyirwaho, bigira ingaruka kuri PCR.
6. 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

Imfashanyigisho:

 Akagari Byoroshye Byoroshye RT-qPCR Kit-Taqman