Hakakino liluwelo noiʻi kāu kiʻina hana a me kā lākou hoʻohana

Hakakino liluwelo noiʻi epekema hula nui i kūlana ma ke kālā no lapaau, forensic epekema, a me ka kālaimeaola. Mahalo i ka hana 'ia he ma i ke ao o ka DNA a me RNA, i ke kanaka mea hiki, e makaikai i ka genome o ka meaola e hoʻoholo i ka causative Agena, e ike i ka makemake nukeleaʻakika i loko o ka läÿau, o nāʻakika, etc.

Hakakino-liluwelo ki ina hana like. He aha ka mea?

Back i loko o nā 70-80s o nā kānakaʻepekema i ka mea mua e decipher i ke kanaka genome. Kēia hanana i impetus i ke kūpono 'ana o ke aaiaoe' enekinia, a me ka hakakino kālaimeaola. Study o ka waiwai o ka DNA a me ka RNA i ke ano o ia mea, ka manawa e hiki ke hoʻohana i kēia mau nukelea nāʻakika no ka mea o ka maʻiʻike, study'ōewe.

E hoomakaukau ai o ka DNA a me ka RNA

Hakakino liluwelo PalekanaEND_LINK ki ina hana like e noi mai ana e hoʻomakaʻia: pinepine keia nukeleaʻakika. Aia i kekahi mau aoao, e koho i kēia mau waiwai, mai na keena o ola meaola. Kēlā i kona pono a me keakea, a me ka mea e manaoia ka wā koho i kekahi ano o ka kaʻawale nukelea nāʻakika i loko o ke gula palapala.

1. e hoomakaukau ai o ka DNA e Marmur. Ka papa hana ninoieo i loko o ka hana maikaʻi i ka hui o ka waiʻona waiwai, precipitates kūpono maemae DNA. Ke downside o keia iaoia o ka hoʻohana 'ana o corrosive waiwai, phenol a me ka chloroform.

2. hoʻokaʻawale o ka DNA ma luna o ka mea hoʻonui. I ka papa kuhikuhiE waiwai i ua hoʻohanaʻaneʻi - ka guanidine thiocyanate (GuSCN). Ua paipai 'ana i ka palapala moʻopane o deoxyribonucleicʻakika i hana kūikawā substrates, mai i ka mea hiki ke a laila ohi ia ma ka mea o ka kūikawā aooa. Eia naʻe GuSCN - mea he inhibitor o ka TCP, a hiki i ka uuku hapa o uaʻia waihoia ma DNA ke ho ololi i ka papa o ka polymerase kaula naoh, i mea nui e hana ana me ka nukelea nāʻakika wā.

3. kimu o impurities. Ka papa hana i kekahi mai ka mua a mākou i loko o ia na nā lātoma lakou iho i ole waiho dehoksiribonukleinovoyʻakika a me impurities. E hana i kēia, e hoʻohana i ka ion poe kuai moni. Ke keakea mea i ole a pau 'ona e hoopau i.

4. Mass loiloi. Kēia papa hana ua hoʻohana 'ia i loko o ka wā hihia oe e ole e pono ai i ka i pololei' ike e pili ana i ka 'ole o ka DNA lātoma, a me ka mea e kiʻi i kekahi mau' ikepili helu hemahema. Ke kumu o ka mea hiki e poino ka mea nukeleaʻakika 'ole ka wā lawelawe detergents, maopopo alkalies.

Mea Maluna o ka noiʻi ki ina hana like

All hakakino-liluwelo ano o ka hana noiʻi, ua māheleʻia i loko o ekolu nui pūʻulu.

1. Amplification (ka hoʻohana 'ana i ka ai iiiaanoaa o nā'ūhū). Kēia nā PCR - polymerase lei naoh, i Radio kūlana nui i loko o na mea he nui o ka PalekanaEND_LINK ki ina hana like.

2. Neamplifikatsionnye. Kēia pūʻulu o nā kiʻina hana a ua pili 'ana i ka hana o ka nukeleaʻakika nā mea huihuina. Ano he kumu hoʻohālike i 3 ano holoi, ma ka situ hybridization, etc.

3. Nā Papahana ka nānā 'ana i ka hoʻomaopopo pono o ka hōʻailona, mai kaʻimi lātoma, ka mea e hoopaa ana i kekahi mau' DNA a RNAʻimi. Laʻana - hybridization'ōnaehana Hybride hoʻopio aku i System pāʻoihana (hc2).

Nā'ūhū i e hoʻohana i ka hakakino liluwelo noiʻi epekema hiki

He nui hakakino PalekanaEND_LINK i kūpono Komo i ka hoʻohana 'ana i ka laulā huahelu o nā'ūhū. Aia ma lalo e hoʻohana 'ia ka hapanui pinepine:

1. kapu'ūhū - "ʻoki" ka DNA lātoma i ka mea e pono ai wahi.

2. DNA polymerase - synthesizes he papalua-ili deoxyribonucleicʻakika lātoma.

3. e hoole transcriptase (e hoole transcriptase) - hoʻohana 'e synthesize DNA, mai ka RNA UAAIIeIA.

4. DNA ligase - He kuleana no ka ho okumu ana o phosphodiester mea paʻa ma waena o nucleotides.

5. exonuclease - e lawe aku oe nucleotides, mai ka pau ana haʻawina o ka deoxyribonucleicʻakika lātoma.

PCR - ka walaʻauʻana ano o ka DNA amplification

Polymerase kaula naoh (PCR) ua hoʻomano hoʻohana 'ia i loko o kēia hakakino kālaimeaola. Kēia hana, ma a he hookahi DNA lātoma hiki ke loaa he nui helu o ke kope (amplifying lātoma).

O ka papa kuhikuhi hana o PCR:

-ʻike, o ka maʻi;

- cloning DNA Hoʻohana,'ōewe.

E lawe mai i ka polymerase kaula naoh pono ke kēia hehee wale: loiloi mua DNA lātoma, he thermostable DNA polymerase (Taq a Pfu), deoxyribonucleotide phosphates (kumu o ka naikokene kumu), na primers (2 primer 1 DNA lātoma) a iho i kekahi aooa?'ōnaehana a e alakai a pau reactions.

PCR ninoieo a ekolu anuu: denaturation, primer annealing a me ka elongation.

1. denaturation. I ka wela o 94-95 kekelē Kelekia ma proshodit uhai i ka hydrogen paa ana ma waena o nā kaula hiloʻelua o ka DNA, a me ka hopena ua loaa elua hookahi-ili nā lātoma.

2. annealed primers. I ka wela o 50-60 degere centigrade ia lŘlŘ ka piʻiʻana primers ma nā wēlau o ka hookahi-ili nukeleaʻakika nā lātoma e like me ke 'ano o ka complementarity.

3. Elongation. I ka wela o ka 72 degere ua synthesized kaikamahine papalua ili deoxyribonucleicʻakika nā lātoma.

DNA sequencing

Hakakino liluwelo noiʻi epekema pinepine koi ike i ke kaʻina o nā nucleotides i loko o ka lātoma o deoxyribonucleicʻakika. E hoʻoholo i ka aaiaoe kivila kaʻina. Hakakino lapaʻau 'o ka wā e hiki mai, e e ma muli o ka ike i loaa ma ka manao paa o ke kanaka ke kaʻina,.

ʻO kēia mau 'ano o ka sequencing:

• sequencing ma ka Maxam-Gilbert;
• Sanger sequencing;
• pyrosequencing;
• nanoporovoe sequencing.