Buccal Swab/FTA Card DNA Isolation Kit Genomic DNA Extraction or Purifiaction Kit mai Buccal Swabs
ʻO ka wehewehe ʻana o ka pahu:
E hoʻomaʻemaʻe koke i ka DNA genomic kiʻekiʻe mai nā laʻana buccal swab/FTA Card.
◮ʻAʻole hoʻohaumia ʻo RNase:ʻO ka DNA-Only Column i hāʻawi ʻia e ka kit e hiki ai ke wehe i ka RNA mai ka DNA genomic me ka hoʻohui ʻole ʻana i ka RNase i ka wā o ka hoʻokolohua, e pale ana i ka hale hana mai ka haumia ʻana e ka exogenous RNase.
◮wikiwiki wikiwiki:ʻOi aku ka hana o Foregene Protease ma mua o nā protease like, a ʻeli koke i nā ʻāpana kiko;maʻalahi ka hana, a hiki ke hoʻopau ʻia ka hana unuhi DNA genomic i loko o 20-80 mau minuke.
◮ʻoluʻolu:Hana ʻia ka centrifugation ma ka lumi wela, a ʻaʻohe pono no ka 4 ° C haʻahaʻa haʻahaʻa centrifugation a i ʻole ethanol precipitation o DNA.
◮Palekana:ʻAʻole koi ʻia ka hoʻoheheʻe ʻana i nā mea hoʻoheheʻe organik.
◮kūlana kiʻekiʻe:ʻO ka DNA genomic i unuhi ʻia he mau ʻāpana nui, ʻaʻohe RNA, ʻaʻohe RNase, a me ka haʻahaʻa haʻahaʻa haʻahaʻa, hiki ke hoʻokō i nā koi o nā hoʻokolohua like ʻole.
◮Pūnaehana micro-elution:Hiki iā ia ke hoʻonui i ka manaʻo o ka genomic DNA, kahi kūpono no ka ʻike ʻana i lalo a i ʻole ka hoʻokolohua.
wehewehe
Hāʻawi kēia kit i kahi ala kūpono a wikiwiki hoʻi e kiʻi ai i ka DNA genomic kiʻekiʻe mai nā swabs buccal a me FTA Card (nā kiko koko).Ke hoʻohana nei i kā mākou hui's DNA-wale nō silica membrane spin kolamu a me ka formula, i hui pū me Foregene Protease, kiʻekiʻe-concentrate, kiʻekiʻe kiʻekiʻe genomic DNA hiki ke unuhi i loko o 80 minuke.ʻO ke kolamu hoʻomaʻemaʻe liʻiliʻi i hoʻolālā ʻia e hoʻopaʻa i ka DNA genomic, a hiki ke hoʻokaʻawale ʻia ka DNA me kahi liʻiliʻi (15μl) ʻōnaehana elution e hoʻonui i ka ʻike o ka DNA genomic i loaʻa, kahi kūpono no ka ʻike ʻana i lalo a i ʻole ka hoʻokolohua.Hiki i ka pahu ke hana i hoʻokahi a ʻoi aku paha nā laʻana i ka manawa, a ʻaʻole pono ke kaʻina hoʻomaʻemaʻe i ka unuhi ʻana o nā mea kūlohelohe e like me phenol, chloroform, a me ka isopropanol a i ʻole ethanol precipitation, a maʻalahi ka hana a mālama manawa.
Nā kikoʻī
50 Hoʻomākaukau
Nā ʻāpana pahu
| Pale ST1 |
| Hoʻopaʻa ST2 |
| Linear Acrylamide |
| Hoopaa PW |
| Hoʻopaʻa WB |
| Hoʻopaʻa EB |
| Foregene Protease |
| DNA-wale wale ke kolamu |
| Nā kuhikuhi |
Nā hiʻohiʻona & pono
-ʻAʻohe RNase contamination: ʻO ka DNA-Only Column i hāʻawi ʻia e ka kit e hiki ai ke wehe i ka RNA mai ka genomic DNA me ka ʻole o ka hoʻohui ʻana i ka RNase i ka wā o ka hoʻokolohua, e pale ana i ka hale hana mai ka haumia ʻana e ka exogenous RNase.
ʻO ka wikiwiki wikiwiki: ʻoi aku ka ʻoi aku o ka ʻoihana Foregene Protease ma mua o nā protease like, e ʻeli koke i nā laʻana;hana maʻalahi.
-Maikaʻi: Hana ʻia ka centrifugation ma ka lumi wela, a ʻaʻohe pono no ka 4 ° C haʻahaʻa haʻahaʻa centrifugation a i ʻole ethanol precipitation o DNA.
- Palekana: ʻAʻole koi ʻia ka hoʻoheheʻe reagent organik.
-Kiʻekiʻe kiʻekiʻe: ʻO ka DNA genomic i unuhi ʻia he mau ʻāpana nui, ʻaʻohe RNA, ʻaʻohe RNase, a me ka haʻahaʻa haʻahaʻa haʻahaʻa, hiki ke hoʻokō i nā koi o nā hoʻokolohua like ʻole.
- Pūnaehana Micro-elution: Hiki iā ia ke hoʻonui i ka ʻike o ka genomic DNA, kahi kūpono no ka ʻike ʻana i lalo a i ʻole ka hoʻokolohua.
Ke noi pahu
He kūpono ia no ka hoʻomaʻemaʻe ʻana i ka DNA genomic mai kēia mau laʻana: buccal swabs, FTA Card (nā koko koko).
Ka mālama ʻana a me ka mālama ʻana
-Hiki ke mālama ʻia kēia kit no 12 mau mahina ma lalo o nā kūlana maloʻo ma ka lumi wela (15-25 ° C);inā pono e mālama ʻia no ka manawa lōʻihi, hiki ke mālama ʻia ma 2-8 ° C.
'Ōlelo Aʻo: Inā mālama ʻia ma kahi wela haʻahaʻa, hiki i ka hāʻina ke hiki i ka ua.Ma mua o ka hoʻohana ʻana, e hōʻoia e waiho i ka hopena i loko o ka pahu i ka lumi wela no kekahi manawa.Inā pono, e hoʻomaʻamaʻa i ka wai ʻauʻau 37 ° C no 10 mau minuke e hoʻoheheʻe i ka wai, a hui ʻia ma mua o ka hoʻohana ʻana.
-Foregene Protease solution he kumu kū hoʻokahi, e hana ana i ka wā e mālama ʻia ai i ka lumi wela no ka manawa lōʻihi (3 mau mahina);ʻoi aku ka maikaʻi o kāna hana a me ka paʻa i ka wā e mālama ʻia ai ma 4 ° C, no laila e ʻōlelo ʻia e mālama iā 4 ° C, e hoʻomanaʻo ʻaʻole e mālama iā ia ma -20 ° C.
Paʻa ke kolamu hoʻomaʻemaʻe
Ma keia kit, i loko o ka genomic DNA extraction hana, ka hoʻomaʻemaʻe kolamu ua pololei adsorbed ma luna o ka hāpana enzymatic lysis hui me ka centrifugation kaʻanuʻu, a me ka hoʻomaʻemaʻe kolamu hiki ke ālai 'ia ma muli o incomplete enzymization a me kiʻekiʻe viscosity o ka hāpana.
ʻO nā kumu kūpono e like me kēia:
1. ʻAʻole i hoʻopau ʻia ka hoʻoheheʻe enzymatic o nā ʻano kiko.
Manaʻo Manaʻo: Hiki ke hoʻonui ʻia ka manawa hana laʻana o Foregene Protease a i ʻole hiki ke lawe ʻia ka supernatant ma hope o ka centrifugation ma 12,000 rpm (~13,400 × g) no 5 min.
2. ʻO ka hoʻohana nui ʻana i nā mea kikoo a i ʻole nā kikoʻī nui.
Manaʻo: ʻOi aku ka maikaʻi ʻaʻole ma mua o 1 Buccal swab i ka hāpana;inā ʻoi aku ka nui o ka hāpana, e hoʻonui i ke ʻano o Buffer ST1, Foregene Protease, hoʻopaʻa ST2 e like me ia.
3. He kiʻekiʻe loa ka laʻana viscosity.
Manaʻo: Hiki ke hoʻoheheʻe ʻia nā laʻana me 10 mM o Tris-HCl ma mua o ka unuhi ʻana i ka DNA genomic.
4. Ua omo ʻia nā ʻāpana o ke kāleka koko.
Manaʻo Manaʻo: Hiki ke hoʻolōʻihi ʻia ka manawa centrifugation transient o ka ʻanuʻu 6 o ke kiko koko (FTA Card).
Haʻahaʻa haʻahaʻa a ʻaʻohe DNA
Nui nā ʻano mea like ʻole e pili ana i ka hua DNA genomic, e like me ke kumu hoʻohālike, nā kūlana mālama laʻana, ka hoʻomākaukau ʻana i ka laʻana, manipulation, etc.
ʻAʻole hiki ke loaʻa ka DNA Genomic i ka wā o ka unuhi ʻana
ʻO nā kumu kūpono e like me kēia:
1. ʻO ka mālama pono ʻole ʻana i nā laʻana a i ʻole mālama ʻia no ka lōʻihi loa e alakaʻi i ka genomic DNA degradation.
Manaʻo Manaʻo: Pono e hoʻāʻo hou ʻia nā swabs waha, a ʻaʻole pono e hoʻohana i nā swabs i mālama ʻia no nā hana unuhi DNA genomic;Pono nā laʻana kiko koko e hōʻoia i ka maikaʻi o ka maikaʻi a ʻaʻole lōʻihi ka manawa mālama.
2. ʻAʻole hiki ke hoʻohana ʻia ka liʻiliʻi o ka ʻiʻo i ka unuhi ʻana o ka DNA genomic pili.
Manaʻo Manaʻo: E hahai i nā ʻōlelo aʻoaʻo no ka hoʻopili ʻana i ka buccal swab sampling i loko o ke alakaʻi hana, a holoi i nā manawa he nui i hiki i hiki ke hoʻopili ʻia nā cell i ka swab waha no ka unuhi ʻana i ka DNA genomic;no ka lawe ʻana i kahi laʻana koko, hiki ke hoʻonui ʻia ka wahi ʻoki koko.
3. ʻAʻole mālama pono ʻia ka Foregene Protease, ka hopena i ka emi ʻana o kāna hana a i ʻole ka hana ʻole.
Manaʻo: E hōʻoia i nā kūlana mālama o ka Foregene Protease a i ʻole e hoʻololi iā ia me kahi Foregene Protease hou no ka hopena enzymatic.
4. He lōʻihi loa ka mālama pono ʻole ʻana o ka pahu a i ʻole ka manawa mālama ʻana, e hopena i ka hemahema o kekahi mau mea i loko o ka pahu.
Manaʻo: E kūʻai i kahi pahu hoʻokaʻawale DNA Buccal swab hou no nā kaʻina hana pili.
5. ʻAʻole hoʻohui ʻo Buffer WB i ka ethanol piha.
Manaʻo Manaʻo: E hōʻoia i ka hoʻohui ʻana o WB i ka leo kūpono o ka ethanol piha.
6. ʻAʻole i hoʻohui pono ʻia ka eluent i ke kiʻi silicone.
Manaʻo Manaʻo: E hoʻohui i ka 65 °C eluent pre-warmed eluent i ka waena o ka membrane silicone a waiho i ka lumi wela no 5 min e hoʻonui i ka pono elution.
Hoʻokaʻawale ʻia ka DNA genomic haʻahaʻa
ʻO nā kumu kūpono e like me kēia:
1. ʻO ka mālama pono ʻole ʻana i nā laʻana a i ʻole mālama ʻia no ka lōʻihi loa e alakaʻi i ka genomic DNA degradation.
Manaʻo Manaʻo: ʻoi aku ka maikaʻi o ka hoʻāʻo hou ʻana i nā swabs waha, a ʻaʻole pono e hoʻohana ʻia nā swabs i mālama ʻia no ka unuhi ʻana i ka DNA genomic.
2. Inā he liʻiliʻi loa ka nui o ka laʻana kiko, e emi ana ka ʻike DNA genomic i unuhi ʻia.
Manaʻo Manaʻo: E hahai i nā ʻōlelo aʻoaʻo laʻana o ka waha i loko o ke alakaʻi hana, e holoi i nā manawa he nui i hiki ke hoʻopili ʻia nā cell i ka swab waha no ka unuhi ʻana i ka DNA genomic.
3. ʻAʻole mālama pono ʻia ka Foregene Protease, ka hopena i ka emi ʻana o kāna hana a i ʻole ka hana ʻole.
Manaʻo: E hōʻoia i nā kūlana mālama o ka Foregene Protease a i ʻole e hoʻololi iā ia me kahi Foregene Protease hou no ka hopena enzymatic.
4. Nā pilikia eluent.
Manaʻo: E hoʻohana i ka Buffer EB no ka elution;inā hoʻohana ʻo ddH2ʻO a i ʻole nā eluent ʻē aʻe, e hōʻoia i ka pH o ka eluate ma waena o 7.0-8.5.
5. ʻAʻole i hoʻokomo pono ʻia ka ʻeluate i ka kulu.
Manaʻo: E hoʻohui i nā kulu eluent i ka waena o ka membrane silicone a waiho i ka lumi wela no 5 min e hoʻonui i ka pono elution.
6. He liʻiliʻi loa ka wai elution.
Manaʻo: E hoʻohana i ka eluent no ka elution DNA genomic e like me ka mea i koi ʻia ma nā kuhikuhi, ʻaʻole liʻiliʻi ma lalo o 15 μl.
Hoʻokaʻawale ʻia ka maʻemaʻe haʻahaʻa o ka DNA genomic
Hiki i ka hoʻomaʻemaʻe DNA genomic haʻahaʻa ke alakaʻi i ka hopena a i ʻole nā hopena maikaʻi ʻole o nā hoʻokolohua o lalo, e like me: ʻaʻole hiki ke ʻoki ʻia nā enzymes, ʻaʻole hiki i ka PCR ke loaʻa i ka ʻāpana gene o ka hoihoi, etc.
ʻO nā kumu kūpono e like me kēia:
1. Heteroprotein pollution, RNA pollution.
Nānā: ʻAʻole holoi ʻia ke kolamu hoʻomaʻemaʻe me ka hoʻohana ʻana i ka Buffer PW;ʻaʻole holoi ʻia ke kolamu hoʻomaʻemaʻe holoi Buffer PW me ka hoʻohana ʻana i ka wikiwiki centrifugal pololei.
Manaʻo: E hōʻoia ʻaʻohe ua i loko o ka supernatant ma mua o ka hoʻohui ʻana i ka ethanol;e holoi i ke kolamu hoʻomaʻemaʻe e like me nā kuhikuhi, a ʻaʻole hiki ke haʻalele ʻia kēia hana.
2. Impurity ion pollution.
Nānā: Ua kāpae ʻia a holoi ʻia paha ke kolamu hoʻomaʻemaʻe ʻo Buffer WB i hoʻokahi wale nō, e hopena i ke koena ionic contamination.
Manaʻo Manaʻo: E holoi pono i ka Buffer WB 2 manawa e like me ke kuhikuhi ʻana e wehe i nā koena ion e like me ka hiki.
3. Hoʻohaumia i ka enzyme RNA.
Nānā: Ua hoʻohui ʻia nā RNases haole i ka pale;ʻAʻole i pololei ka holoi ʻana i ka Buffer PW, e hopena i nā koena RNase, e pili ana i nā hana hoʻokolohua RNA i lalo, e like me ka transcription in vitro.
Manaʻo Manaʻo: Foregene series nucleic acid isolation kits hiki ke wehe i ka RNA me ka ʻole o ka hoʻohui ʻana o RNase, no laila ʻaʻole pono e hoʻohui i ka RNase Kāleka Swab/FTA Card DNA Isolation Kit;e hahai i nā kuhikuhi no ka kolamu holoi holoi holoi Buffer PW, a ʻaʻole hiki ke haʻalele ʻia kēia hana.
4. Ke koena ethanol.
ʻIkepili: ʻAʻole i hana ʻo Buffer WB i ka centrifugation paipu ʻole ma hope o ka holoi ʻana i ke kolamu hoʻomaʻemaʻe.
Manaʻo Manaʻo: E hana i ka hana centrifugation paipu hakahaka kūpono e like me nā kuhikuhi.
5. ʻO nā mea haumia ʻē aʻe.
Ka nānā 'ana: 'A'ole mālama 'ia nā la'ana i mālama 'ia a i 'ole nā la'ana kūikawā.
Manaʻo: E mālama mua i ka hāpana e like me ke kuhikuhi ʻana.
