Njira maselo kwachilengedwenso kafukufuku ndi ntchito

Maselo kwachilengedwenso njira kafukufuku mbali yofunika kwambiri mu mankhwala amakono, sayansi azamalamulo, ndi zamoyo. Ndiyamika patsogolo kuphunzira DNA ndi RNA, munthu amatha kufufuza matupi a chamoyo kudziwa wothandizila causative, kuzindikira kufunika asidi mkati mwa osakaniza zidulo, etc.

Maselo-kwachilengedwenso njira. Ndi chiyani icho?

Back mu 70-80s asayansi anali oyamba kumvetsa matupi a anthu. chochitika linapereka kuti chitukuko cha zomangamanga majini ndi zamoyo. Kuphunzira katundu wa DNA ndi RNA kwatanthauza kuti tsopano n'zotheka ntchito zidulo izi mkati cholinga cha matenda matenda, kuphunzira mbewu.

Kukonzekera DNA ndi RNA

Maselo kwachilengedwenso njira yodziwira zimafunika kuyambira zakuthupi: kawirikawiri izi asidi mkati. Pali njira zingapo kusankha zinthu izi kwa maselo a zinthu zamoyo. Aliyense ali ndi ubwino ndi kuipa kwake, ndipo tiyenera kuziganizira posankha njira kusakonda zidulo mkati mwa mawonekedwe woyera.

1. Kukonzekera DNA ndi Marmur. njira tichipeza pochiza osakaniza mowa zinthu, precipitates chifukwa DNA koyera. The downside njira ndi ntchito zinthu zikuwononga, phenol ndi chloroform.

2. kudzipatula DNA pa Boom lapansi. Thunthu waukulu ntchito pano - ndi guanidine thiocyanate (GuSCN). Iwo amachita mafunsidwe a deoxyribonucleic acid pa magawo apadera, amene akhoza kenako anasonkhana mwa gawo lotetezedwa wapadera. Komabe GuSCN - ndi woletsa wa TCP, ndipo ngakhale gawo laling'ono la kamakhala waikamo mu DNA kumakhudza zochita za polymerase unyolo anachita, chimene chiri chofunika pamene akugwira zidulo mkati.

3. chamvula zosafunika. njira isiyana ndi anthu yoyamba kuti mamolekyulu okha si waikamo asidi dehoksiribonukleinovoy ndi zosafunika. Kuchita izi, ntchito ion kuzizira. The sangathe ndiye kuti zinthu zonse akhoza kuthetsa.

4. Misa mosamala. Njira ntchito milandu pamene simuyenera kukhala ndi mfundo zolondola zokhudza Kapangidwe ka DNA, ndi kufunika kutenga ziwerengero zotsatirazi. Chifukwa chake kuti mkati kapangidwe asidi zikhoza kuonongeka pogwiritsa detergents, makamaka alkalies.

Gulu la njira zofufuzira

Onse njira maselo-zamoyo kafukufuku anawagawa m'magulu atatu aakulu:

1. amveke (ntchito kuchuluka kwa michere). Izi zikuphatikizapo PCR - polymerase unyolo anachita, amene amagwira ntchito yofunika kwambiri mu njira zimene matenda.

2. Neamplifikatsionnye. Gulu la njira amagwirizana mwachindunji ndi ntchito zosakaniza mkati acid. Zitsanzo ndi 3 mitundu zinawafutapo, mu situ mwakusakaniza, etc.

3. Njira zochokera kuzindikira chizindikiro pa kafukufuku molekyulu, amene chimango ndi yeniyeni DNA kapena RNA kafukufuku. Mwachitsanzo - mwakusakaniza dongosolo Hybride Jambulani System njira (hc2).

Michere kuti angagwiritsidwe ntchito njira maselo kwachilengedwenso kafukufuku

Njira zambiri maselo matenda zimagwiritsa ntchito zosiyanasiyana waukulu michere. Pansipa ntchito zambiri:

1. chiletso enzyme - "kudula" ndi DNA ziwalo zofunika.

2. DNA polymerase - synthesizes wachiphamaso-Shackleton deoxyribonucleic asidi molekyulu.

3. n'zosiyana transcriptase (chigamulocho transcriptase) - ntchito lithe DNA kuchokera Chinsinsi RNA.

4. DNA ligase - ndi udindo mapangidwe kumangidwa phosphodiester pakati nucleotides.

5. exonuclease - amachotsa nucleotides ku mbali mapeto a deoxyribonucleic asidi molekyulu.

PCR - njira yaikulu ya DNA amveke

Polymerase unyolo anachita (PCR) chimagwiritsidwa ntchito mu sayansi amakono maselo. njira imeneyi, imene umodzi DNA akhoza zopezera chiwerengero chachikulu cha magazini (amplifying molekyulu).

Ntchito yaikulu ya PCR:

- matenda a matenda;

- cloning DNA zigawo, Gene.

Kuti akachite polymerase unyolo anachita amafuna zinthu zotsatirazi: koyamba DNA, ndi thermostable DNA polymerase (Taq kapena Pfu), deoxyribonucleotide phosphates (magwero a zapansi asafe), ndi primers (2 phunziroli 1 DNA) ndi lokha dongosolo gawo lotetezedwa amene angafunikire kuchititsa zochita zonse.

PCR imakhala njira zitatuzi: denaturation, phunziroli annealing ndi elongation.

1. denaturation. Pa kutentha 94-95 madigiri Celsius proshodit kuswa mgwirizano wa hydrogen pakati maunyolo awiri a DNA, ndi chifukwa ife tikufika awiri mamolekyulu osakwatiwa-Shackleton.

2. annealed primers. Pa kutentha madigiri 50-60 centigrade amapezeka primers inalowa malekezero a mamolekyulu osakwatiwa-Shackleton mkati asidi monga mtundu wa mapolojekiti.

3. Elongation. Pa kutentha madigiri 72 apanga mwana wamkazi awiri Shackleton mamolekyulu deoxyribonucleic acid.

DNA molumikiza

Maselo njira kafukufuku kwachilengedwenso zambiri amafuna kudziwa za ndondomeko ya nucleotides mu molekyulu asidi deoxyribonucleic. Kudziwa malamulo opanga khalidwe anafufuza. diagnostics maselo a m'tsogolo kuzikidwa kudziwa tingaphunzire mu mtima wa akuyambira anthu.

Zotsatirazi mitundu ya molumikiza:

• molumikiza ndi Maxam-Gilbert;
• Sanger molumikiza;
• pyrosequencing;
• nanoporovoe molumikiza.