Pehea ka nui o kou ike

E pili ana iā Nka ʻili ʻana o ka ʻakika ucleic

ʻO ka hoʻomaka a me ka hopena o ka waikawa nucleic i hoʻomaʻemaʻe ʻia

Paʻakikī nā mea a pau i ka hoʻomaka ʻana, ʻoi aku ka nui o ka waika nucleic hoʻomaʻemaʻe

Hoʻomaka i nā hoʻokolohua molekula, no nā hoʻokolohua ma hope

He hopena koʻikoʻi ka holomua a i ʻole ka hāʻule ʻole.

ʻO Trace/ultra-trace UV spectrophotometer ka mea i makemake nui ʻia e nā mea noiʻi ʻepekema no ka mea hiki iā ia ke ʻike i ka neʻe ʻana o ka waikawa nucleic a me nā koena ion a me ka paʻakai maʻalahi, wikiwiki a me ka waiwai.

E like me kā mākou e ʻike ai, ʻo A260 ke ʻano o ka waiwai nucleic acid absorbance OD, ʻo A280 ke ʻano o ka protein absorbance absorbance OD value, A230 ke ʻano o ka paʻakai paʻakai absorbance absorbance OD waiwai, no laila hiki iā A260 ke hoʻololi i ka wai nucleic acid, A260/280 ʻo ia ke koena protein, A260/230 ʻo ia ke koena o ka paʻakai paʻakai, akā ʻo ka nui o nā hopena i ʻike ʻia e nā mea noiʻi.kuʻuna"e manaʻoʻiʻo hiki iā ia ke wehewehe i ka maʻemaʻe o DNA a me RNA ikahi ākea.ʻAʻole ia wale nō ka mea maʻamau no ka ʻike ʻana i ka hua nucleic acid a me ka maʻemaʻe, hoʻohana wale ʻia ia ma ke ʻano he kuhikuhi, a ʻaʻole ia he "pae gula".

Hoʻomaopopo ka manual Thermo Fisher ND-2000 i ka hilinaʻi o nā hopena hōʻike

ʻO ke kumu, ʻo nā hopena i loaʻa ma ka hoʻohana ʻana i ka micro/ultramicro UV spectrophotometer e hōʻike wale ana i ka hua a me ka maʻemaʻe o ka waika nucleic mai ka ʻaoʻao, a he nui nā mea hoʻohālikelike (ala optical, sample volume, sample concentration, pH value, other ions e pili ana i ke ana absorbance, etc.) , ʻaʻole pololei ka helu OD i ana ʻia.I ka manawa like, ma muli o ka nele o ka kikoʻī o ka waiwai hoʻopaʻa ʻana i ka waiwai, ka DNA i hoʻopaʻa ʻia, ka DNA pālua, RNA, dNTPs, a me kekahi mau protein i loaʻa nā puʻupuʻu absorption ma 260nm hawewe lōʻihi, a ʻo ka manaʻo i hoʻoholo ʻia e A260 wale nō ʻaʻole pono ka DNA maoli a i ʻole RNA.ʻAʻole hiki ke noʻonoʻo ʻia ka noʻonoʻo ʻana, a me kona kūpaʻa a me ka degradation.

No laila pehea e hoʻoholo ai i ka maikaʻi o kā mākou nucleic acid?Ma waho aʻe o ka hoʻohana ʻana i ka micro/ultramicro UV spectrophotometer no ka ʻike ʻana, koi ʻia nā ʻano e like me ka electrophoresis gel agarose e hōʻike maka i ka maʻemaʻe a me ka pono o ka nucleic acid, a e hōʻike i ka ʻike ʻana i kekahi ʻano.Ma kēia ala, hiki ke hilinaʻi ʻia ka hua nucleic acid a me ka maʻemaʻe mai nā hopena ʻike o nā ʻano like ʻole.

Hoʻohālikelike pakuhi hoʻokolohua

Ua unuhi ʻia ʻo Genomic DNA o nā pūnaewele H1299 me ka hoʻohana ʻana i nā kits Extraction DNA o ka hui A, a ua ʻike ʻia ka haumia koʻikoʻi, ka hopena i nā waiwai OD kiʻekiʻe.

(Ka waiwai ana o OD a me ka electropherogram pili)

Helu helu loiloi

Aia he "maʻamau gula" no ka hua nucleic acid a me ka hoʻāʻo maikaʻi?

E like me kā mākou ʻike, ʻaʻohe ala maʻalahi, wikiwiki a maʻalahi.Inā he spectrophotometer, electrophoresis gel, a i ʻole spectrometry nuipa, hōʻike lākou a pau i ka ʻike a me ka maʻemaʻe o ka waikawa nucleic i loko o ka hopena mai nā ʻano like ʻole, a pono lākou e hoʻokolokolo ʻia ma ke kuhikuhi ʻana i kekahi i kekahi.

ʻO ka papa kuhikuhi loiloi maikaʻi loa i nā manawa a pauka noi hoʻokolohua hahai.ʻAʻole pili ia i ka maikaʻi o ka hoʻohuli ʻana a me ka PCR amplification.ʻO ka loaʻa ʻana o nā huahana hoʻonui hilinaʻi a me nā waiwai Ct, a loaʻa i kahi kaʻina holoʻokoʻa ma ke kaʻina ʻana he kūleʻa ka hoʻoneʻe nucleic acid.

No ka hōʻuluʻulu ʻana, pono e hoʻohālikelike ʻia ka manaʻo o ke kumumanaʻo ratio-only e ka manaʻo o "ka hoʻohana ʻana i nā hopena hoʻokolohua maikaʻi i lalo e like me nā ʻāpana ʻoki maikaʻi"!

Manaʻo ʻia ʻo Foregene DNA RNA kime e loaʻa ai ka maʻemaʻe DNA/RNA kiʻekiʻe:

https://www.foreivd.com/reagent/dna-isolation-series/

https://www.foreivd.com/reagent/rna-isolation-series/