Hababka baarista bayoolojiga kelli iyo isticmaalkooda

Kelli noolaha habab cilmi ka ciyaaraan door muhiim ah ka daawo casri ah, sayniska foreensiga, iyo biology. Thanks to horumarka waxbarasho ee DNA iyo RNA, qofka waa inay awoodaan inay sahamiyaan hiddawadayaasha ee u shaqeeyo loo ogaado wakiilka sababaya, in la aqoonsado nucleic acid la doonayo in isku dar ah ee acids, iwm

Hababka kelli-noolaha. Waa maxay?

Back ee 70-80s ah aqoonyahaniinta ahaa kii ugu horeeyay ee rayi hiddawadayaasha aadanaha. Dhacdadani waxa ay dar dar geliyay horumarinta injineernimada hidaha iyo biology kelli. Study of sifooyinka DNA iyo RNA ayaa loogu tala galay in ay hadda waa suurto gal in la isticmaalo kuwaas oo acids nucleic ujeeddo ah ogaanshaha cudurka, hiddo-waxbarasho.

Diyaarinta DNA iyo RNA

noolaha kelli hababka ogaanshaha baahan bilaabin wax: inta badan this nucleic acid. Waxaa jira siyaabo dhowr ah in ay doortaan, kuwaas oo walxaha laga unugyada noolaha. Kasta wuxuu leeyahay faa'iidooyinka iyo dhibaatooyinka ay, waana in la tixgeliyaa marka aad dooranayso hab ka mid ah la takooro acids nucleic qaab saafi ah.

1. Diyaarinta DNA by Marmur. Habka wuxuu ka kooban yahay daaweynta isku dar ah ee walxaha khamriga, precipitates keentay DNA saafi ah. Dhinaca kale ee habkan waa isticmaalka walxaha xoqe, phenol iyo chloroform.

2. Go'doon ee DNA on kaca ah. walax ugu muhiimsan ee ah in waxaa halkan loogu isticmaalayaa - waa thiocyanate guanidine (GuSCN). Waxay kor u dhigaalka ee deoxyribonucleic acid on muhashada gaarka ah, kaas oo waxaa la soo ururiyey danbe karaa iyadoo qofka ku xiranyahay gaar ah. Si kastaba ha ahaatee GuSCN - waa inhibitor ah TCP ah, iyo xitaa qayb yar oo ah in uu dhigo DNA saamayn ku yeelan karaan koorsada cadaanyo silsilad polymerase ah, taas oo muhiim ah marka la acids nucleic shaqeeya.

3. Da'itaan ee nijaasta ahaa. Habka ka duwan kuwii hore in in u taagoo ee laftooda aan wax shubo acid dehoksiribonukleinovoy iyo nijaasta ahaa. Si arrintan loo sameeyo, u isticmaal exchangers ra'iyi ah. Faa'iida waa in walxaha dhan ma degi karaan.

4. Baaritaanka Mass. Habkani waxaa loo isticmaalaa in kiisaska marka aad uma baahna in ay leeyihiin macluumaad sax ah oo ku saabsan qaab dhismeedka of Britain DNA ah, iyo baahida loo qabo in la helo tirakoob qaar ka mid ah. Sababtuna waxay tahay, in qaab dhismeedka acid nucleic dhaawacmi karaa marka saabuun, gaar ahaan alkalies.

Qoondaynta habab cilmi

All hababka kelli-noolaha ee cilmi-baarista waxaa lagu qaybiyey saddex kooxood oo waaweyn:

1. weyneeye (iyadoo la isticmaalayo hannaanka ah ensaymes). Tan waxaa ka mid PCR - cadaanyo silsilad polymerase, kaas oo kaalin muhiim ah in badan oo ka mid ah hababka lagu ogaanayo.

2. Neamplifikatsionnye. Kooxdaan hababka si toos ah oo la xidhiidha shaqo ee isku acid nucleic. Tusaale ahaan waa waskhaamiin 3 nooc, in hybridization situ, iwm

3. Hababka ku salaysan aqoonsiga signal ka soo Britain ayaa baaritaan, kaas oo xidhana in DNA ama RNA baaritaan gaar ah. Tusaale - Nidaamka hybridization xal System Qabsashada Hybride (HC2).

Ensaymes in waxa loo isticmaali karaa habab kelli cilmi noolaha

farsamooyin badan oo lagu ogaanayo kelli ku lug isticmaalka kala duwan ee ensaymes. Hoos waxaa ku qoran waxaa inta badan la isticmaalo:

1. enzyme xadaynta - "gooyaa" Britain DNA qaybaha lagama maarmaanka ah.

2. DNA polymerase - daydaa Britain acid a double-ku xayiran deoxyribonucleic.

3. transcriptase noqonaynin (beddeli transcriptase) - loo isticmaalaa in synthesize DNA ka template RNA ah.

4. ligase DNA - ayaa mas'uul ka ah formation of bonds phosphodiester dhexeeya nucleotides.

5. exonuclease - saaraysaa nucleotides ka qaybo dhammaadka Britain ee acid deoxyribonucleic ah.

PCR - habka aasaasiga ah ee cod-weyneeye DNA

silsilad polymerase reaction (PCR) waxaa si balaadhan looga isticmaalaa in biology kelli casriga ah. Habkani, taas oo Britain ah DNA hal laga heli karaa tiro badan oo nuqullo (Britain jaha).

Shaqooyinka ugu muhiimsan ee PCR:

- ogaanshaha cudurrada;

- cloning qaybaha DNA, Gene.

Si ay u fuliyaan reaction silsilad polymerase u baahan yahay arrimaha soo socda: Britain DNA hore, a DNA polymerase thermostable (Taq ama Pfu), phosphates deoxyribonucleotide (ilaha saldhig nitrogen), primers (2 Primer 1 DNA Britain) iyo laftiisa nidaamka kayd ah oo ay sameyn karaan oo dhan reactions.

PCR ka kooban tahay saddex tallaabo: denaturation, annealing Primer iyo muruqa.

1. denaturation. At heerkul ah 94-95 degrees Celsius proshodit jebin bonds hydrogen dhexeeya laba silsiladood oo ah DNA ah, iyo iyada oo sababtu tahay aan helno laba taagoo hal-ku xayiran.

2. primers annealed. At heerkul ah 50-60 degrees Centigrade dhacdaa primers goysiga at darafyadiisa oo u taagoo acid nucleic hal-ku xayiran sida ay nooca complementarity.

3. muruqa. At heerkul ah 72 darajo waa gabadhii ku dhexsameeysmo double ku xayiran taagoo acid deoxyribonucleic.

horumarintooda DNA

Hababka kelli cilmi noolaha inta badan u baahan tahay aqoon isku xigxiga ee nucleotides ee Britain ayaa ah deoxyribonucleic acid. Si loo go'aamiyo ka code genetic sequenced. Baadhista xanuunada kelli ee mustaqbalka waxaa lagu salayn doonaa aqoonta qaadatay go'aan ka soo baxay isku xigxiga ee aadanaha.

The noocyada soo socda ee horumarintooda:

• horumarintooda by the-Maxam Gilbert;
• Sanger horumarintooda,
• pyrosequencing;
• nanoporovoe horumarintooda.