ʻO Kina ʻoihana maʻalahi a wikiwiki hoʻi i ka DNA/Rna One-Step Probe Rt-Qpcr Kit
Ke manaʻo nei kā mākou paʻa e hana pono, e lawelawe ana i kā mākou poʻe kūʻai a pau, a me ka hana ʻana i ka ʻenehana hou a me ka mīkini hou no ka Professional China Convenient and Quick DNA/Rna One-Step Probe Rt-Qpcr Kit, Ke hana nei kā mākou paʻa me ke kaʻina hana o ka "kūpono-ma muli o ka hui ʻana, hana ʻia nā kānaka, lanakila-lanakila hui".Manaʻolana mākou hiki iā mākou ke loaʻa kahi pilina aloha maikaʻi me ka mea kālepa mai nā wahi a puni.
Manaʻo kā mākou ʻoihana e hana pono, lawelawe i kā mākou poʻe kūʻai āpau, a me ka hana mau ʻana i ka ʻenehana hou a me ka mīkini hou.Kina Taq DNA Polymerase a me Qpcr, Hoʻolālāʻia kā mākou mauʻoihana a me nā kaʻina hana e hōʻoia i ka loaʻaʻana o kā mākou mau mea kūʻai i ka laulā o nā huahana a me nā hoʻonā me nā laina manawa hāʻawi pōkole loa.Loaʻa ʻia kēia hoʻokō e kā mākou hui akamai a ʻike.Ke ʻimi nei mākou i nā poʻe makemake e ulu pū me mākou a puni ka honua a kū i waho o ka lehulehu.I kēia manawa, loaʻa iā mākou ka poʻe e apo ana i ka lā ʻapōpō, loaʻa ka ʻike, ke aloha hoʻopololei i ko lākou mau manaʻo a hele i kahi mamao ma mua o ka mea a lākou i manaʻo ai he hiki ke loaʻa.
Nā wehewehe kime
ʻO ka 2X Real PCR maʻalahiTMHāʻawi ʻia ʻo Mix-Taqman e ka Real Time PCR EasyTM-ʻO ka pahu Taqman kahi ʻōnaehana premix hou e hoʻohana ana i nā ʻōkuhi fluorescent kūikawā no ka Real Time PCR amplification reactions, hiki ke hoʻomaikaʻi nui i ka kikoʻī o ka huahana a me ka naʻau pane.Hāʻawi ʻia ʻo ROX ma ke ʻano he kala hoʻomalu kūloko.
2X PCR maʻalahiTMLoaʻa i ka Mix-Taqman ka Taq DNA Polymerase kūikawā o Foregene.Ke hoʻohālikelike ʻia me nā enzyme Taq maʻamau, loaʻa iā ia nā pōmaikaʻi o ka hoʻonui hoʻonui kiʻekiʻe, ka mana amplification kikoʻī ikaika a me ka haʻahaʻa haʻahaʻa haʻahaʻa.Hiki iā ia ke hōʻemi i ka hoʻonui ʻole kikoʻī a hoʻomaikaʻi i ka pololei o PCR.
Nā kikoʻī
Maʻalahi PCR manawa maoliTM-Taqman | ||||
ʻO ka hui pū ʻana (20μl system) | QP-01021 | QP-01022 | QP-01023 | QP-01024 |
200T | 500T | 1000T | 2000T | |
2× Maʻalahi PCR maoliTMHui-Taqman | 1 ml × 2 | 1.7 ml × 3 | 1.7 ml × 6 | 1.7 ml × 12 |
20× ROX Peʻa Waikane | 200 μl | 0.5 ml | 1 ml | 1 ml × 2 |
Dnase-Free ddH2O | 1.7 ml | 1.7 ml × 2 | 10 ml | 20 ml |
Aʻo | 1 | 1 | 1 | 1 |
Nā hiʻohiʻona & pono
■ Maʻalahi—2X PCR Mix e hōʻemi i ka hewa hoʻokolohua a me ka manawa hana
■ Hiki ke pale aku i ka hoʻonui ʻole a me ka hoʻokumu ʻana o ka dimer kikoʻī kikoʻī—ʻoi aku ka maikaʻi a me ka hoʻomaka wela Taq enzyme.
■ Kiʻekiʻe sensitivity—hiki ke ʻike i nā kope haʻahaʻa o ka template
■ Maikaʻi maʻalahi—kūpono me ka hapa nui o nā mea kani PCR i ka manawa maoli
Ke noi pahu
ka nānā ʻana qPCR
Ka holo hana
Kiʻi kiʻi
Ka mālama ʻana a me ka mālama ʻana
Pono e mālama ʻia kēia pahu mai ka mālamalama a mālama ʻia ma -20 ℃.Inā hoʻohana pinepine ʻia, hiki ke mālama ʻia ma 4 ℃ no kahi manawa pōkole (10 mau lā).
Ke manaʻo nei kā mākou paʻa e hana pono, e lawelawe ana i kā mākou poʻe kūʻai a pau, a me ka hana ʻana i ka ʻenehana hou a me ka mīkini hou no ka Professional China Convenient and Quick DNA/Rna One-Step Probe Rt-Qpcr Kit, Ke hana nei kā mākou paʻa me ke kaʻina hana o ka "kūpono-ma muli o ka hui ʻana, hana ʻia nā kānaka, lanakila-lanakila hui".Manaʻolana mākou hiki iā mākou ke loaʻa kahi pilina aloha maikaʻi me ka mea kālepa mai nā wahi a puni.
Kina ʻoihanaKina Taq DNA Polymerase a me Qpcr, Hoʻolālāʻia kā mākou mauʻoihana a me nā kaʻina hana e hōʻoia i ka loaʻaʻana o kā mākou mau mea kūʻai i ka laulā o nā huahana a me nā hoʻonā me nā laina manawa hāʻawi pōkole loa.Loaʻa ʻia kēia hoʻokō e kā mākou hui akamai a ʻike.Ke ʻimi nei mākou i nā poʻe makemake e ulu pū me mākou a puni ka honua a kū i waho o ka lehulehu.I kēia manawa, loaʻa iā mākou ka poʻe e apo ana i ka lā ʻapōpō, loaʻa ka ʻike, ke aloha hoʻopololei i ko lākou mau manaʻo a hele i kahi mamao ma mua o ka mea a lākou i manaʻo ai he hiki ke loaʻa.
ʻAʻohe hōʻailona hoʻonui
1. Nalo ka hana o ka Taq DNA Polymerase i loko o ka pahu ma muli o ka mālama pono ʻole a i ʻole ka pau ʻana o ka pahu.
Manaʻo: E hōʻoia i nā kūlana mālama o ka pahu;e hoʻohui hou i ka nui kūpono o Taq DNA Polymerase i ka ʻōnaehana PCR a i ʻole e kūʻai i kahi PCR Kikoʻo Real Time hou no nā hoʻokolohua pili.
2. Aia ka nui o nā mea pale o Taq DNA Polymerase i loko o ka DNA template.
Manaʻo Manaʻo: E hoʻomaʻemaʻe hou i ka template a i ʻole e hōʻemi i ka nui o ka template i hoʻohana ʻia.
3. ʻAʻole kūpono ka Mg2 + kaʻina.
Manaʻo Manaʻo: ʻO ka Mg2+ ka ʻike o ka 2 × Real PCR Mix a mākou e hāʻawi ai he 3.5mM.Eia nō naʻe, no kekahi mau kumu mua a me nā laʻana, ʻoi aku ka kiʻekiʻe o ka Mg2+.No laila, hiki iā ʻoe ke hoʻohui pololei iā MgCl2 no ka hoʻonui ʻana i ka ʻike Mg2+.Manaʻo ʻia e hoʻonui i ka Mg2+ 0.5mM i kēlā me kēia manawa no ka optimization.
4. ʻAʻole kūpono nā kūlana PCR amplification, a ʻaʻole kūpono ke kaʻina kumu a i ʻole ka ʻike.
Manaʻo: hōʻoia i ka pololei o ke kaʻina kumu mua a ʻaʻole i hoʻohaʻahaʻa ʻia ka primer;inā ʻaʻole maikaʻi ka hōʻailona hoʻonui, e hoʻāʻo e hoʻohaʻahaʻa i ka mahana annealing a hoʻoponopono pono i ka ʻike kumu.
5. He liʻiliʻi a ʻoi aku ka nui o ka template.
Manaʻo Manaʻo: E hana i ka hoʻoheheʻe ʻana i ka gradient linearization template, a koho i ka manaʻo kumu me ka hopena PCR maikaʻi loa no ka hoʻokolohua PCR ʻOiaʻiʻo.
He kiʻekiʻe loa ka waiwai fluorescence o ka NTC
1.Reagent contamination i ka wā o ka hana.
Manaʻo: Hoʻololi me nā mea hou no nā hoʻokolohua PCR ʻOiaʻiʻo.
2. Ua loaʻa ka contamination i ka wā o ka hoʻomākaukau ʻana i ka ʻōnaehana hopena PCR.
Manaʻo Manaʻo: E lawe i nā mea pale pono i ka wā e hana ai, e like me: ke komo ʻana i nā mīkina lima latex, me ka hoʻohana ʻana i ka piko pipette me kahi kānana, etc.
3. Ua hoʻohaʻahaʻaʻia nā kumu mua, aʻo ka hoʻohaʻahaʻaʻana o nā kumu mua e hoʻonui i ka hoʻonuiʻole.
Manaʻo Manaʻo: E hoʻohana i ka electrophoresis SDS-PAGE e ʻike inā ua hoʻohaʻahaʻa ʻia nā primers, a hoʻololi iā lākou me nā primers hou no nā hoʻokolohua PCR Real Time.
Primer dimer a i ʻole ka hoʻonui kikoʻī ʻole
1. ʻAʻole kūpono ka Mg2 + kaʻina.
Manaʻo Manaʻo: ʻO ka Mg2 + ka nui o ka 2 × Real PCR EasyTM Mix a mākou e hāʻawi ai he 3.5 mM.Eia nō naʻe, no kekahi mau kumu mua a me nā laʻana, ʻoi aku ka kiʻekiʻe o ka Mg2+.No laila, hiki iā ʻoe ke hoʻohui pololei iā MgCl2 no ka hoʻonui ʻana i ka ʻike Mg2+.Manaʻo ʻia e hoʻonui i ka Mg2+ 0.5mM i kēlā me kēia manawa no ka optimization.
2. He haʻahaʻa loa ka mahana annealing PCR.
Manaʻo Manaʻo: E hoʻonui i ka mahana o ka PCR annealing e 1 ℃ a i ʻole 2 ℃ i kēlā me kēia manawa.
3. He lōʻihi loa ka huahana PCR.
Manaʻo Manaʻo: Pono ka lōʻihi o ka huahana PCR Real Time ma waena o 100-150bp, ʻaʻole ʻoi aku ma mua o 500bp.
4. Ua hoʻohaʻahaʻaʻia nā kumu mua, aʻo ka hoʻohaʻahaʻaʻana o nā kumu mua e alakaʻi i keʻano o ka amplification kiko'ī.
Manaʻo Manaʻo: E hoʻohana i ka electrophoresis SDS-PAGE e ʻike inā ua hoʻohaʻahaʻa ʻia nā primers, a hoʻololi iā lākou me nā primers hou no nā hoʻokolohua PCR Real Time.
5. ʻAʻole kūpono ka ʻōnaehana PCR, a i ʻole ka liʻiliʻi o ka ʻōnaehana.
Manaʻo Manaʻo: He ʻuʻuku loa ka ʻōnaehana pane PCR e hōʻemi i ka pololei o ka ʻike.ʻOi aku ka maikaʻi o ka hoʻohana ʻana i ka ʻōnaehana pane i ʻōlelo ʻia e ka mea hana PCR quantitative e holo hou i ka hoʻokolohua PCR Real Time.
ʻAʻole hiki ke hoʻihoʻi hou i nā waiwai quantitative
1. Ua hewa ka mea kani.
Manaʻo Manaʻo: Loaʻa paha nā hewa ma waena o kēlā me kēia puka PCR o ka mea kani, e hopena i ka maikaʻi ʻole o ka hana hou ʻana i ka wā o ka mālama ʻana a i ʻole ka ʻike ʻana.E ʻoluʻolu e nānā e like me nā kuhikuhi o ka mea hana pili.
2. ʻAʻole maikaʻi ka maʻemaʻe hāpana.
Manaʻo Manaʻo: E alakaʻi nā laʻana haumia i ka hana hou ʻana o ka hoʻokolohua, ʻo ia hoʻi ka maʻemaʻe o ka template a me nā kumu mua.ʻOi aku ka maikaʻi o ka hoʻomaʻemaʻe hou ʻana i ka template, a hoʻomaʻemaʻe maikaʻi ʻia nā primers e SDS-PAGE.
3. He lōʻihi loa ka hoʻomākaukau ʻana o ka ʻōnaehana PCR a me ka manawa mālama.
Manaʻo Manaʻo: E hoʻohana i ka ʻōnaehana PCR ʻOiaʻiʻo no ka hoʻokolohua PCR ma hope koke o ka hoʻomākaukau ʻana, a mai haʻalele iā ia no ka lōʻihi loa.
4. ʻAʻole kūpono nā kūlana PCR amplification, a ʻaʻole kūpono ke kaʻina kumu a i ʻole ka ʻike.
Manaʻo: hōʻoia i ka pololei o ke kaʻina kumu mua a ʻaʻole i hoʻohaʻahaʻa ʻia ka primer;inā ʻaʻole maikaʻi ka hōʻailona hoʻonui, e hoʻāʻo e hoʻohaʻahaʻa i ka mahana annealing a hoʻoponopono pono i ka ʻike kumu.
5. ʻAʻole kūpono ka ʻōnaehana PCR, a i ʻole ka liʻiliʻi o ka ʻōnaehana.
Manaʻo Manaʻo: He ʻuʻuku loa ka ʻōnaehana pane PCR e hōʻemi i ka pololei o ka ʻike.ʻOi aku ka maikaʻi o ka hoʻohana ʻana i ka ʻōnaehana pane i ʻōlelo ʻia e ka mea hana PCR quantitative e holo hou i ka hoʻokolohua PCR Real Time.