Aia kekahi mauʻokoʻa likeʻole e hiki ke hoʻohanaʻia no ka nānāʻana a me ke kiʻiʻana i ka DNA ma hope o ka hoʻokaʻawaleʻiaʻana o nā mea e ka electrophoresis gel. Ma waena o nā koho he nui,ʻo kēia mauʻelimaʻelima ka hoʻomaka mua loa me ka Ethidium bromide, kahi i hoʻohana nuiʻia. I ka hanaʻana me kēia kaʻina hana, he mea nui kaʻikeʻole i kaʻokoʻa ma waena o nā maʻi wale nō akā i nā pōʻaiapili ola kino.
01'Ohidium Bromide
Hiki paha i ka Ethidium bromide ka mea i kaulana loa i ka nānāʻana i ka DNA. Hiki ke hoʻohanaʻia kēia waiʻai i loko o ka paila gel, ma ka pahukū koho a iʻole eʻoki i ka gel ma hope o ka holoʻana. Hoʻopili nā molecules o ke kinopa i nā kaula DNA a me ka hawanawana ma lalo o ke kukui Uila, e hōʻike ana iāʻoe kahi i paʻa ai nā apo i loko o ka pahu. Ma mua o kona pōmaikaʻi,ʻo ka lalo ma lalo,ʻo ka ethidium bromide he kinohu kūpono, no laila pono e mālama ponoʻia me ka mālama nui. Ma muli o nā pilikino olakino kūpono o ka hanaʻana me kēia gel, ua loaʻa nā mea'ē aʻe. 02 Kakau Gold
Hiki ke hoʻohanaʻia i ka diawera kelepona gula e hoʻokaʻawale i nā DNAʻelua a iʻole keʻano hoʻokahi, aiʻole e pale i ka RNA. ʻO SYBR Gold kekahi o nā meaʻokoʻa'ē aʻe i ka ethidium bromide e pāʻani i ka mākeʻai a ua manaʻoʻia heʻoi aku ka maikaʻi ma mua o ka bumide ethidium. Hiki i kaʻili ke hāʻawi aku i ka UV o ka fluorescence he 1000 ke kiʻekiʻe i ka manawa e kauʻia ana i nā meaola nucleic. A laila, komo i loko o kaʻeleʻele lahilahi a me ka piʻi nui o nā'āpana gels a hiki ke hoʻohanaʻia i ka formaldehyde gels. Ma muli o keʻano haʻahaʻa o ka'ōmole o ka mole laulā,ʻaʻole i koiʻia. Ua hoʻomohala a kūʻaiʻia nā mea e paʻa ana i nā mea e pili ana i nā mea i loaʻa iā ia (mai ka hoʻomakaʻana o SYBR Gold) i ka SYBR Safe a me SYBR Green iʻoi aku ka maluhia o nā mea'ē aʻe i ka ethidium bromide.
03 SYBR Green
Hoʻomākaukauʻia ka SYBR Green I a me II (hou, kūʻaiʻia e Molecular Probes) no nā kumu likeʻole. Ma muli o ko lākou nakinakiʻana i DNA, ua manaʻoʻia lākou he mau mutagens a no ka mea, pono e mālamaʻia me ka mālama. SyBR Green Uaʻoi aku koʻu maʻalahi no ka hoʻohanaʻana me DNA i hoʻopiliʻiaʻelua,ʻoiaiʻo SyBR Green II, ma kekahiʻaoʻao'ē aʻe,ʻoi aku ka maikaʻi no ka hoʻohanaʻana me DNA a hoʻokahi RNA. E like me keʻano o kaʻulu Ethidium Bromide i kaulana,ʻo kēia mau mea pale i keʻano o ke kukui ma lalo o ke kukui UV. 04 I ka hale mālama
Hoʻonohonohoʻiaʻo SyBR Safe e lilo i meaʻoluʻolu palekana i ka Ethidium Bromide a me nā mea'ē aʻe SYBR. ʻAʻole ia i manaʻoʻia he lapalapa weliweli a hiki ke hoʻohanaʻia ma o nā pūnaewele hoʻohaʻahaʻa maʻamau (ʻo ia hoʻi i lalo o ke kahawai), no ka mea,ʻo ka hōʻailona o ka hoʻokolohua e hōʻike anaʻaʻole he lāʻauʻawaʻawa. Hōʻike ka hōʻoia he nui aʻaʻoleʻole he genotoxicity ma nā pūnaewele Hamster Embryo (SHE), nā lymphocytes kanaka, nā lymphoma kiole a me ka hōʻike AMES. Hiki ke hoʻohanaʻia kaʻili me ka poli polū e hoʻomāmā ai i kaʻikeʻana o ka DNA a hāʻawi aku i ka maikaʻiʻoi aku no ka hoʻopiliʻana aku. 05 Eva Green
ʻO Eva Green he'ōmaʻomaʻu'ōmaʻomaʻo'ōmaʻomaʻo i loaʻa i ka paleʻana i ka PCR i kahi liʻiliʻi ma mua o nā mea'ē aʻe. ʻO kēia ka mea maikaʻi loa no nā noi e like me ka PCR-manawa maoli. He mea maikaʻi nō hoʻi ināʻoe e hoʻohana ana i nā gels haʻahaʻa haʻahaʻa no ka hoʻopakeleʻana i ka DNA. He paʻa loa ia i nā mahana wela a he haʻahaʻa haʻahaʻa konaʻano pono'ī, akā, he loloa maikaʻi loa ke kauʻia i ka DNA. Hōʻikeʻiaʻo Eva Green i ka haʻahaʻa ikiʻole aʻaʻoleʻole i ka cytotoxicity.