Akụkọ - mmalite teknụzụ mmịpụta nucleic acid

ⅠNucleic acidimmalite

Nucleic acid na-ekewa n'ime deoxyribonucleic acid (DNA) na ribonucleic acid (RNA), n'etiti nke RNA kewara na ribosomal RNA (rRNA), messenger RNA (mRNA) na nyefee RNA (tRNA) dị ka ọrụ ya dị iche iche.A na-etinyekarị DNA na nucleus, mitochondria na chloroform, ebe RNA na-ekesa na cytoplasm.Dị ka ihe ndabere akụrụngwa nke mkpụrụ ndụ ihe nketa, mmịpụta nucleic acid na-arụ ọrụ dị oke mkpa na nyocha ihe ọmụmụ ihe omimi na nyocha nke ụlọ ọgwụ.Ntinye uche na ịdị ọcha nke mmịpụta nucleic acid ga-emetụta PCR na-esote, usoro, nrụpụta vector, mgbaze enzyme na nnwale ndị ọzọ.

Ⅱ Nucleic acid mmịpụta na usoro dị ọcha

① Usoro mmịpụta phenol/chloroform

Mwepụta phenol/chloroform bụ usoro oge gboo maka mmịpụta DNA, nke na-ejikarị ihe mgbaze organic abụọ dị iche iche na-agwọ ihe nlele ahụ, na-agbaze DNA nke sitere na acid nucleic na mmiri mmiri, lipids na akụkụ organic, yana protein n'etiti akụkụ abụọ ahụ.Usoro a nwere uru nke ọnụ ala dị ala, ịdị ọcha dị elu na mmetụta dị mma.Ihe ọghọm ndị ahụ bụ ọrụ mgbagwoju anya na ogologo oge.

② Usoro Trizol

Usoro Trizol bụ usoro oge gboo maka mmịpụta RNA.A na-ekewa usoro Trizol n'ime mmiri mmiri na akụkụ organic mgbe centrifugation na chloroform, nke RNA na-agbaze na mmiri mmiri, a na-ebufe mmiri mmiri na tube ọhụrụ EP, a na-enweta mmiri ozuzo mgbe agbakwunyere isopropanol, wee mee ka ethanol dị ọcha.Usoro a dabara adaba maka mmịpụta RNA site na anụ ahụ anụmanụ, sel na nje bacteria.

③ Usoro nchacha kọlụm Centrifugal

Centrifuge kọlụm ọcha usoro nwere ike adsorb DNA kpọmkwem site pụrụ iche silicon matrix adsorption ihe, mgbe RNA na protein nwere ike ịgafe were were, na mgbe ahụ na-eji elu nnu ala PH ikpokọta nucleic acid, obere nnu elu PH uru elution kewapụ na sachaa nucleic acid.Uru ndị dị na ya bụ itinye uche dị ọcha dị elu, nkwụsi ike dị elu, ọ dịghị mkpa maka ihe mgbaze organic, na ọnụ ala dị ala.Ọdịmma ya bụ na ọ dị mkpa ka ọ bụrụ centrifud nzọụkwụ site nzọụkwụ, usoro ọrụ ndị ọzọ.

④ Usoro bead magnetik

Usoro ihe ndọta magnetik bụ iji kewaa mkpụrụ ndụ anụ ahụ site na lysate, hapụ acid nucleic dị na nlele ahụ, mgbe ahụ, a na-etinye mkpụrụ ndụ nucleic acid n'elu oghere magnetik, ebe adịghị ọcha dị ka protein na sugars na-ahapụ. mmiri mmiri.Site na steepụ nke cell anụ ahụ kewara, magnetik bead ejikọta na nucleic acid, nucleic acid ịsacha, nucleic acid elution, wdg, a na-emecha nweta nucleic acid dị ọcha.Uru ndị dị mfe na-arụ ọrụ na obere oge iji, na-enweghị mkpa nke nzọụkwụ centrifugation.Ọ nwere ala teknuzu chọrọ na ike ịghọta akpaka na uka ọrụ.Ngwakọta a kapịrị ọnụ nke magnetik bead na nucleic acid na-eme ka acid nucleic amịpụtara nwee nnukwu itinye uche na ịdị ọcha.Mwepu bụ na ọnụ ahịa ahịa dị ugbu a dị oke ọnụ.

⑤ Ụzọ ndị ọzọ

Na mgbakwunye na ụzọ anọ ndị dị n'elu, enwere mgbawa na-esi ísì ụtọ, usoro nnu na-etinye uche, usoro anionic detergent, usoro ultrasonic na usoro enzymatic, wdg.

Ⅲ Ụdị mmịpụta nucleic acid

Foregene nwere ikpo okwu PCR Direct Direct nke ụwa, kọlụm abụọ RNA Isolation platform(DNA-naanị + RNA naanị).Ngwaahịa ndị bụ isi gụnyere DNA/RNA kewapụ ngwa, PCR na Direct PCR reagents usoro nyocha ụlọ nyocha.

① Mkpokọta mmịpụta RNA

Ngụkọta ihe nlele RNA gụnyere ọbara, mkpụrụ ndụ, anụ ahụ anụmanụ, osisi, nje, wdg. Enwere ike nweta ịdị ọcha dị elu na mkpokọta RNA zuru oke site na mmịpụta RNA zuru oke, nke enwere ike iji na RT-PCR, nyocha mgbawa, ntụgharị in vitro, molecular cloning, Dot Blot na nnwale ndị ọzọ.

metụtara mbụNgwa iche iche RNA

Ngwakọta Mkpokọta RNA anụmanụ--Ngwa ngwa na ịrụ ọrụ nke ọma wepụta mkpokọta RNA ịdị ọcha na ịdị elu dị elu site na anụ ahụ anụmanụ dị iche iche.

Ngwakọta Mkpokọta Cell RNA--Enwere ike nweta mkpokọta RNA sachachara nke ọma na nke dị elu site na sel dị iche iche emebere n'ime nkeji iri na otu.

Ngwakọta Mkpokọta ihe ọkụkụ RNA--Wepụ mkpokọta RNA dị elu ngwa ngwa site na nlele osisi nwere obere polysaccharide na polyphenol.

Ngwa nkewapụ RNA Viral--Wepụ ngwa ngwa ma sachaa nje virus RNA site na nlele dịka plasma, serum, mmiri ahụ na-enweghị sel na ihe ndị na-eme omenala sel.

② mmịpụta DNA nke mkpụrụ ndụ ihe nketa

Genomic DNA mmịpụta samples gụnyere ala, feces, ọbara, mkpụrụ ndụ, anụmanụ anụ ahụ, osisi, nje, wdg Genomic DNA mmịpụta nwere ike ji mee ihe na enzyme mgbaze, DNA Library ewu, PCR, antibody nkwadebe, Western blot hybridization analysis, gene chip, high. -usoro ihe omume na nnwale ndị ọzọ.

metụtara mbụNgwa iche iche DNA

Ngwa anụ ahụ DNA dịpụrụ adịpụ--Mwepụta ngwa ngwa na ịdị ọcha nke DNA genomic site na isi mmalite dị iche iche, dị ka anụ ahụ anụmanụ, mkpụrụ ndụ, wdg.

Ọbara DNA Midi Kit (1-5ml)--Sachaa ngwa ngwa genomic DNA dị elu site na ọbara anticoagulated (1-5ml).

Buccal Swab/FTA Kaadị DNA kewapụrụ--Sachaa ngwa ngwa genomic DNA dị elu site na nlele kaadị swab/FTA.

Ngwa nkewapụta DNA osisi--Sachapụ ngwa ngwa wee nweta DNA genomic dị elu site na nlele osisi (gụnyere polysaccharides na ihe atụ osisi polyphenol)

③ Mwepụta nke Plasmid

Plasmid bụ ụdị DNA obere molekul okirikiri dị n'ime sel, nke bụ ihe na-ebukarị DNA recombination.Usoro nke mmịpụta plasmid bụ iwepụ RNA, kewapụ plasmid na DNA genomic nje bacteria, wepụ protein na adịghị ọcha ndị ọzọ iji nweta plasmid dị ọcha.

General Plasmid Obere Kit--Sachapụ ngwa ngwa plasmid DNA dị elu site na nje bacteria gbanwere maka nyocha ihe ọmụmụ ihe ndụ dị ka mgbanwe na mgbaze enzyme.

④ Ụdị mmịpụta ndị ọzọ, mmịpụta miRNA, wdg.