Ngwa ngwa ngwa ngwa PCR Kit-Taqman
Nkọwa ngwa:
◮Usoro nkwalite PCR pụrụ iche na-eme 2 × QuickEasyTMReal PCR Mix-Taqman dakọtara karịa.
◮Hot-start Foregene HS Taq Polymerase nwere arụmọrụ nkwalite dị elu, mmetụta mmụba dị elu, yana nkọwa nkwalite dị elu.
◮2 × Ọ dị mfeTMReal PCR Mix -Taqman na-eji sistemu pụrụ iche nke Foregene kwalitere iji kwalite nghọta na nkọwapụta nke nchọpụta usoro usoro, nke enwere ike iji maka mkpụrụ ndụ ihe nketa na detuo mkpebi mgbanwe ọnụọgụ nọmba wee nweta nsonaazụ ziri ezi.
◮Ngwaahịa a na-abịa na ROX n'ime ntụ ntụ ntụ, nke enwere ike iji wepụ mgbaama ndabere na njehie mgbaàmà n'etiti olulu mmiri, nke dị mma maka ndị ahịa iji na ụdị PCR dị iche iche.
Nkọwa
2 × QuickEasyTMReal PCR Mix-Taqman nyere site na QuickEasyTMNgwa PCR Real Time Kit-Taqman bụ usoro ngwakọta nna ukwu nke ọgbọ ọhụrụ emebere maka mmeghachi nkwalite PCR Real Time site na iji nyocha fluorescent akọwapụtara.Isi ya Foregene HS Taq DNA Polymerase dabere na Mgbanwe Antibody, ejiri ya na sistemụ PCR nke Foregene na-emeziwanye, nwere nkọwapụta dị elu na arụmọrụ nkwalite.N'iji ya na ngwakọta PCR nkịtị, ọ nwere ikike mmụba siri ike yana ọnụego adabaghị.Enwere ike iji ya na mmeghachi omume PCR nke fluorescence quantitative iji belata mmụba na-abụghị nke akọwapụtara yana kwalite izi ezi nke PCR.Ọ nwere ike melite nkọwapụta ngwaahịa yana mmetụta mmeghachi omume.N'otu oge ahụ, a na-enye ROX ka ọ bụrụ ihe ntụnye aka n'ime.
The QuickEasyTMReal Time PCR Kit-Taqman kit nwere a sara mbara akporo ngwa na ego nke template, na ike nweta a ezi ọkọlọtọ usoro na dị iche iche quantitative mpaghara, na ike n'ụzọ ziri ezi quantify na chọpụta lekwasịrị mkpụrụ ndụ ihe nketa na ezi repeatability na elu obi ike.
Nkọwapụta
200 nkeji,500 ụzọ,1000 nkwadebe,2000 Preps
Ngwa ngwa
| Ọ dị mfeTMReal Time PCR Kit-Taqman(20μlusoro) | ||||
| Ngwa ọdịnaya | QP-01121 | QP-01122 | QP-01123 | QP-01124 |
| 200 Preps | 500 nkwadebe | 1000 Preps | 2000 Preps | |
| 2×Ngwa ngwaMfeTM Ezigbo PCR Ngwakọta- Taqman | 1 ml×2 | 1.7 ml×3 | 1.7 ml×6 | 1.7 ml×12 |
| 20×ROX ntụaka agbaji | 200μL | 0,5 ml | 1 ml | 1 ml×2 |
| ddH na-efu DNAase2O | 1.7 ml | 1.7 ml×2 | 10 ml | 20 ml |
| IFU | 1ibe | 1ibe | 1ibe | 1ibe |
Atụmatụ & uru
■ Usoro nkwalite PCR pụrụ iche na-eme 2 × QuickEasyTMReal PCR Mix-Taqman dakọtara karịa.
■ Hot-start Foregene HS Taq Polymerase nwere nrụmọrụ nkwalite dị elu, mmetụta mgbago elu dị elu, yana nkọwa nkwalite dị elu.
■ 2× Ọ dị mfeTM Real PCR Mix -Taqman na-eji sistemu pụrụ iche nke Foregene kwalitere iji kwalite nghọta na nkọwapụta nke nchọpụta usoro usoro, nke enwere ike iji maka mkpụrụ ndụ ihe nketa na detuo mkpebi mgbanwe ọnụọgụ nọmba wee nweta nsonaazụ ziri ezi.
■ Ngwaahịa a na-abịa na ROX n'ime ntụ ntụ ntụ, nke enwere ike iji wepụ mgbanaka ndabere na njehie n'etiti olulu mmiri, nke dị mma maka ndị ahịa iji na ụdị PCR dị iche iche.
Ngwa ngwa
■ Fluorescence quantitative PCR maka ndebiri quantitative analysis;
■ Cnkwalite PCR nke mbụ;
■ Enwere ike iji ya mee nchọpụta allele.
Nchekwa na Shelf ndụ
Ekwesịrị ịchekwa ngwa ahụ na -20 Celsius C n'ọchịchịrị;Ọ bụrụ na a na-ejikarị ya eme ihe, a pụkwara ịchekwa ya na 4 Celsius maka obere oge (jiri ya n'ime ụbọchị 10).
Real Time PCR primer ụkpụrụ imewe
Na-aga n'ihu Primer na Reverse Primer
Maka Real Time PCR, imewe primer dị ezigbo mkpa.Primers metụtara nkọwapụta na arụmọrụ nke nkwalite PCR, enwere ike hazie ya na ntụnyere ụkpụrụ ndị a:
Ogologo oge mbụ: 18-30bp.
Ọdịnaya GC: 40-60%.
Uru Tm: Akụrụngwa imewe nke mbụ, dị ka Primer 5, nwere ike inye uru Tm nke primer.Ụkpụrụ Tm nke ndị na-ebuli elu na nke mgbada kwesịrị ịdị nso dịka o kwere mee.A nwekwara ike iji usoro mgbako Tm: Tm = 4 °C (G + C) + 2 Celsius C (A + T).Mgbe ị na-arụ PCR, a na-ahọrọ okpomọkụ dị n'okpuru uru Tm primer nke 5 Celsius dị ka okpomọkụ na-ekpo ọkụ (mmụba kwekọrọ na okpomọkụ na-ekpo ọkụ nwere ike ịbawanye nkọwa nke mmeghachi omume PCR).
Ngwaahịa mbụ na PCR:
Design primer PCR amplification ogologo ngwaahịa bụ ọkachamma 100-150bp.
Ekwesịrị ịzere primers imewe na mpaghara nhazi nke abụọ nke ndebiri dị ka o kwere mee.
Zenarị nhazi ntọala 2 ma ọ bụ karịa n'etiti nsọtụ 3 nke elu na ala ala.
Primer 3′ terminal base enweghị ike ịnọ ya na 3 mgbakwunye G ma ọ bụ C.
Ndị primers n'onwe ha enweghị ike ịnwe ihe mgbakwunye, ma ọ bụghị ya, a ga-emepụta usoro ntutu isi, na-emetụta mmụba PCR.
Ekwesịrị kesaa ATCG n'otu n'otu dịka o kwere mee n'usoro nke mbụ, yana ebe njedebe 3′ kwesịrị izere ka T.
Ihe mgbakwunye 1: Mkpọ mgbakwunye akụrụngwa Direct RT-qPCR Kit
1.Cell Lysis Solution
|
| |||
| Ngwa ngwa (24-well lysis sistem / ọma) | DRT-01011-A1 | DRT-01011-A2 | |
| 100 T | 500 T | ||
| AkụkụI | Ihe nchekwa CL | 20 ml | 100 ml |
| Foregene Protease Plus II | 400 μl | 1 ml × 2 | |
| Ihe nchekwa ST | 1 ml × 2 | 10 ml | |
| AkụkụII | Nchicha DNA | 400 μl | 1 ml × 2 |
Ngwakọta 2.RT
|
| |
| Ngwa ngwa (Sistemụ mmeghachi omume 20 μl) | DRT-01011-B1 |
| 200 T | |
| Ngwakọta 5 × Direct RT Mix | 800 μl |
| ddH na-efu RNase2O | 1.7 ml × 2 |
|
| ||
| Ngwa ngwa (Sistemụ mmeghachi omume 20 μl) | DRT-01011-C1 | DRT-01011-C2 |
| 200 T | 1000 T | |
| 2× Direct qPCR Mix-SYBR | 1 ml × 2 | 1.7 ml × 6 |
| 50× ROX ntụaka agbaji | 40 μl | 200 μl |
| ddH na-efu RNase2O | 1.7 ml | 10 ml |
