ʻO ka Novel Coronavirus (SARS-CoV-2) Pākuʻi PCR Manawa maoli
Loaʻa kēia pahu PCR maʻalahi a mākaukau e hoʻohana i ka format lyophilized (ka hana maloʻo maloʻo) no ka mālama lōʻihi.Hiki ke lawe ʻia a mālama ʻia ka pahu ma ka lumi wela a paʻa no hoʻokahi makahiki.Loaʻa i kēlā me kēia paipu o premix nā reagents a pau e pono ai no ka hoʻonui ʻana i ka PCR, me ka Reverse-transcriptase, Taq polymerase, primers, probes, a me nā substrate dNTPs.Pono e hoʻohui i 13ul wai distilled a me 5ul extracted RNA template, a laila hiki ke holo a hoʻonui ʻia ma nā mea kani PCR.
Pono ka mīkini qPCR e hoʻokō i kēia mau koi:
1. E hoʻokomo i ka nui o ka paipu PCR 8 strip 0.2 ml
2. Loaʻa i ʻehā mau ala ʻike maka:
| Kanal | Ka hauʻoli (nm) | Hoʻokuʻu (nm) | Nā Wai i hoʻopaʻa ʻia ma mua |
| 1. | 470 | 525 | FAM, SYBR Green I |
| 2 | 523 | 564 | VIC, HEX, TET, JOE |
| 3. | 571 | 621 | ROX, TEXAS-ULAULA |
| 4 | 630 | 670 | CY5 |
Nā papahana PCR:
7500 Pūnaehana PCR manawa maoli, Biorad CF96, iCycler iQ™ Pūnaehana ʻIke PCR manawa maoli, Stratagene Mx3000P, Mx3005P
ʻO ka paʻakikī o ka lawe ʻana i ke kaulahao anu o Novel Coronavirus nucleic acid detection reagent
Ke lawe ʻia nā reagents ʻike nucleic acid ma kahi lōʻihi, ʻo ka (-20 ± 5) ℃ ka mālama ʻana i ke kaulahao anuanu a me ka lawe ʻana e koi ʻia e hōʻoia i ka bioactive o ka enzyme i loko o nā reagents.No ka hōʻoia i ka hiki ʻana o ka mahana i ka maʻamau, pono ʻia he mau kilokani hau maloʻo no kēlā me kēia pahu o ka nucleic acid hoʻāʻo reagent ʻoi aku ka liʻiliʻi ma mua o 50g, akā hiki ke mau no ʻelua a ʻekolu mau lā.Ma ka manaʻo o ka hana ʻoihana, ʻo ke kaumaha maoli o nā reagents i hoʻopuka ʻia e nā mea hana ma lalo o 10% (a i ʻole ka liʻiliʻi ma mua o kēia waiwai) o ka ipu.ʻO ka hapa nui o ke kaumaha mai ka hau maloʻo, nā pahu hau a me nā pahu pahu, no laila ke kiʻekiʻe loa ke kumukūʻai.
I Malaki 2020, ua hoʻomaka ʻo COVID-19 i kahi ākea nui ma waho, a ua piʻi nui ka noi no ka Novel Coronavirus nucleic acid detection reagent.ʻOiai ke kumukūʻai kiʻekiʻe o ka lawe ʻana aku i nā reagents i ke kaulahao anu, hiki i ka hapa nui o nā mea hana ke ʻae iā ia ma muli o ka nui a me ka loaʻa kālā kiʻekiʻe.
Eia nō naʻe, me ka hoʻomaikaʻi ʻana i nā kulekele hoʻopuka aupuni no nā huahana anti-pandemic, a me ka hoʻonui ʻana i ka mana aupuni ma luna o ke kahe o ka poʻe a me ka logistic, aia ka hoʻonui a me ka maopopo ʻole i ka manawa o ka lawe ʻana i nā reagents, i hopena i nā pilikia huahana koʻikoʻi. e ka halihali ana.ʻO ka lōʻihi o ka lawe ʻana i ka manawa (he mea maʻamau ka manawa o ka lawe ʻana ma kahi o ka hapalua mahina) alakaʻi i ka hāʻule pinepine ʻana o ka huahana ke hiki aku ka huahana i ka mea kūʻai.Ua pilikia kēia i ka hapa nui o nā ʻoihana hoʻoiho nucleic acid reagents.
Ua kōkua ka ʻenehana Lyophilized no ka PCR reagent i ka lawe ʻana i ka Novel Coronavirus nucleic acid detection reagent ma ka honua holoʻokoʻa.
Hiki ke lawe ʻia a mālama ʻia nā lyophilized PCR reagents i ka lumi lumi, ʻaʻole hiki ke hōʻemi wale i ke kumukūʻai o ka lawe ʻana, akā pale pū nō hoʻi i nā pilikia maikaʻi i hoʻokumu ʻia e ke kaʻina kaʻa.No laila, ʻo ka lyophilizing ka reagent ke ala maikaʻi loa e hoʻoponopono ai i ka pilikia o ka lawe ʻana aku i waho.
Hoʻopili ka Lyophilization i ka hoʻoheheʻe ʻana i kahi hopena i loko o kahi kūlana paʻa, a laila hoʻohaʻahaʻa a hoʻokaʻawale i ka mahu wai ma lalo o ke ʻano ʻūhā.Noho ka solute maloʻo i loko o ka pahu me ka hana like a me ka hana.Hoʻohālikelike ʻia me nā reagents wai maʻamau, ʻo ka piha piha lyophilized Novel Coronavirus nucleic acid detection reagent i hana ʻia e Liming Bio i kēia mau hiʻohiʻona:
Paʻa wela ikaika loa:hiki iā ia me ka mālama ʻana ma 56 ℃ no 60 mau lā, a ʻaʻole i loli ka morphology a me ka hana o ka reagent.
Waihona wela maʻamau a me ka lawe ʻana:ʻaʻohe pono o ke kaulahao anu, ʻaʻohe pono e mālama i kahi haʻahaʻa haʻahaʻa ma mua o ka wehe ʻana, hoʻokuʻu piha i ka wahi mālama anu.
Mākaukau-e hoʻohana:lyophilizing o nā mea a pau, ʻaʻohe pono no ka hoʻonohonoho ʻana i ka ʻōnaehana, pale i ka nalowale o nā ʻāpana me ka viscosity kiʻekiʻe e like me ka enzyme.
Nā pahuhopu lehulehu i hoʻokahi paipu:ʻO ka pahuhopu ʻike e uhi ana i ka moʻo coronavirus ORF1ab gene, N gene, S gene e pale aku i ka genovariation virus.I mea e hōʻemi ai i ka hewa hewa, hoʻohana ʻia ke kanaka RNase P gene ma ke ʻano he mana kūloko, i mea e hoʻokō ai i ka pono lapaʻau no ka hoʻokele maikaʻi ʻana.
