Hanyar kwayoyin halitta na bincike da kuma amfani da su

Kwayoyin halittu da bincike hanyoyin taka muhimmiyar rawa a zamani magani, na bincike kimiyya, da kuma ilmin halitta. Godiya ga ci gaba a fannin nazarin DNA da RNA, mutum zai iya nazarin kwayar halittar kwayar halitta, gano mai cutar da cutar, ya gane da ake bukata nucleic acid a cikin cakuda acid, da dai sauransu.

Hanyar kwayoyin halitta-hanyoyin nazarin halittu. Mene ne?

Back a cikin 70-80s daga cikin masana kimiyya ne na farko da decipher da mutum genome. Wannan taron ya ba da hankali ga ci gaba da aikin injiniya da kwayoyin halitta. Binciken dukiya na DNA da RNA ya haifar da gaskiyar cewa yanzu yana yiwuwa a yi amfani da wadannan kwayoyin halittu domin gane da cutar, don nazarin kwayoyin.

Shirin DNA da RNA

Kwayoyin halittu bincike hanyoyin buƙaci fara abu: sau da yawa wannan nucleic acid. Akwai hanyoyi da dama don ware waɗannan abubuwa daga kwayoyin halittu. Kowannensu yana da amfani da rashin amfani, kuma dole ne a la'akari da haka yayin zabar hanyar kawar da kwayoyin nucleic a cikin tsabta.

1. Shirya DNA bisa ga Marmur. Hanyar tana kunshe da maganin cakuda abubuwa tare da barasa, sakamakon sakamakon tsabtaccen DNA. Rashin haɓaka wannan hanya shine amfani da abubuwa masu m: phenol da chloroform.

2. Halittar DNA ta hanyar Boom. Abinda ake amfani dasu shine guanidine thiocyanate (GuSCN). Yana inganta cikin shaida na deoxyribonucleic acid a kan qware substrates, daga abin da shi za a iya baya tattara ta wajen wani musamman buffer. Duk da haka, GuSCN mai hanawa ne na PTC, har ma da wani ɓangare na ɓangaren, wanda aka kama a cikin DNA da aka sata, zai iya rinjayar tafarkin musayar polymerase, wanda ke taka muhimmiyar rawa a aikin tare da acid nucleic.

3. Tsarin tsabta. Hanyar ta bambanta da wadanda suka gabata a cikin cewa kwayoyin deoxyribonucleic sun haɓaka da su, amma marasa tsabta. Don yin wannan, amfani da masu musayar wuta. Rashin haɓaka shi ne cewa ba duk abubuwa zasu iya janyewa ba.

4. Nano zane. Ana amfani da wannan hanya a waɗannan lokuta idan ba a buƙatar bayani game da abun da ke ƙunshe da kwayar halittar DNA ba, amma ya zama dole don samun wasu bayanan lissafi. Wannan ya bayyana cewa gaskiyar kwayoyin nucleic za a iya lalacewa ta hanyar maganin cututtuka, musamman, tare da alkalis.

Ƙayyade na hanyoyin bincike

Dukkan hanyoyin nazarin hanyoyin kwayoyin halitta sun kasu kashi uku:

1. Mahimmanci (ta amfani da wasu enzymes). Wannan ya hada da tsarin PCR-polymerase, wanda ke taka muhimmiyar rawa a cikin yawan hanyoyin bincike.

2. Ba a lakafta ba. Wannan rukuni na hanyoyi suna da alaka da aikin haɗuwa da kwayoyin nucleic acid. Misalan su ne nau'i uku na blotting, daidaitaccen wuri, da dai sauransu.

3. Hanyar da aka danganci karɓar siginar daga wata kwayar bincike wadda ta ɗaura zuwa wani bincike na DNA ko RNA. Misali shi ne tsarin samfurori a cikin tsarin samfurin Hybride (hc2).

Enzymes da za a iya amfani dashi a hanyoyin kwayoyin halitta

Hanyoyi da yawa na tantance kwayoyin halitta sun haɗa da yin amfani da fannoni daban-daban na enzymes. Da ke ƙasa an fi amfani dasu:

1. Ƙuntatawa - "yanke" kwayar halittar DNA cikin sassa masu dacewa.

2. DNA polymerase - haɗakar da kwayoyin halitta biyu na deoxyribonucleic acid.

3. Gyara bayanan (revertase) - ana amfani dashi don hada DNA akan matakan RNA.

4. DNA ligase - alhakin samuwar haɗin phosphodiester tsakanin nucleotides.

5. Gabatarwa - ta kawar da nucleotides daga sassan ƙarshen kwayoyin deoxyribonucleic acid.

PCR ita ce babbar hanya ta kara DNA

Polymerase sarkar dauki (PCR) ne yadu a yi amfani da zamani ilmin sanin kwayoyin halittu. Wannan hanya ce wadda za'a iya samun adadi mai yawa daga kwayoyin halittar DNA guda ɗaya (ƙara yawan kwayoyin halitta).

Babban ayyukan PCR shine:

- Sanin asalin cututtuka;

Cloning DNA, kwayoyin.

Halin musayar polymerase yana buƙatar abubuwa masu zuwa: asali na DNA, DNA polymerase wanda aka samo asali (Taq ko Pfu), da magunguna (2 alamomi ta DNA mole) da kuma tsarin buffer kanta, wanda zai yiwu a gudanar da shi Daga dukkan halayen.

PCR ya ƙunshi matakai uku: denaturation, annealing of primers da elongation.

1. Damawa. A zafin jiki na digirin Celsius 94-95, ƙaddamar da jigilar hydrogen tsakanin sassan DNA guda biyu ya ci gaba, kuma a sakamakon haka muna samun nau'i guda guda biyu.

2. Haɗewa na mahimmanci. A yawan zafin jiki na Celsius na 50-60, an saka alamomi a iyakar ƙwayoyin kwayoyin halittu guda guda guda daya ta hanyar nau'in haɓakawa.

3. Elongation. A yawan zafin jiki na digiri 72, ana kiran labaran 'yar ƙwayoyin halitta biyu na deoxyribonucleic acid.

DNA Sequencing

Tsarin halitta na kwayoyin halitta yana bukatar sanin ilimin nucleotides a cikin kwayar deoxyribonucleic acid. Don sanin da kwayoyin code jerin. Magungunan kwayoyin halitta na nan gaba za su dogara ne akan ilimin da aka samu a ƙayyade jerin mutum.

Ana rarrabe wadannan nau'o'in sukurori masu zuwa:

• Takaddamar da Maxam-Gilbert;
• Sanquire ta yanke shawara;
• Tsarin zane-zane;
• Hanyar da ake kira Nanoporous.