Ukuzalwa kwePCR
PCR (Polymerase Chain Reaction)
Sele ingaphezulu kweminyaka engama-30 ukususela ekuvelisweni kwe-polymerase chain reaction.Ngaphezulu kweminyaka engama-30, emva kokuba abaphengululi abaninzi kwihlabathi jikelele beqhubeka bexhasa kwaye bephucula, itekhnoloji ye-PCR iye yaba yeyona ndlela ixhaphakileyo nesetyenziswa rhoqo kwaye eyona ndlela ibalulekileyo yophando olusisiseko kuyo yonke inkalo yeeSayensi zoBomi.
I-PCR ye-TouchDown, i-Real-Time PCR, i-Multi PCR, njl. njl
Iziphene zeteknoloji ye-PCR yemveli
Ukwahlula i-nucleic acid eyinkimbinkimbi kunyeukukhutshwa:
★ Iteknoloji yePCR yeSintu: iyafuneka
★ PCR ephuma iteknoloji: efunekayo
★ Iisampulu ze-DNA kunye ne-RNA: umahluko omkhulu, iimfuno zokusebenza ezinzima
• Iingozi zomzimba: ii-reagents eziyityhefu ziwonakalisa umzimba
Itekhnoloji ye-PCR yesiNtu kunye netekhnoloji ephuma kwi-prerequisite-nucleic acid yokwahlula kunye nokucocwa.
Nayiphi na iisampulu yebhayoloji kufuneka idlule kuthotho lwesampulu enzima kunye nedinayo yokusetyenzwa kwesampulu ukufumana iisampulu ze-nucleic acid ezihlangabezana neemfuno zobuchwepheshe be-PCR.
Ukwahlula kunye nokutsalwa kwe-DNA kunye ne-RNA ibisoloko ingumsebenzi osisiseko ekufuneka abaphandi besayensi abafanelekileyo bawuphindaphinde yonke imihla.
Ngenxa yomahluko omkhulu phakathi kweesampulu, ukwahlukana kunye neenkqubo zokutsalwa kwe-DNA kunye ne-RNA nazo zahluke kakhulu.Lo msebenzi ufuna inqanaba eliphezulu lobuchule bobugcisa kubaqhubi.Ukwahlula ngokwemveli kunye neendlela zokukhupha zifuna ukudibana kwexesha elide kunye nezinye ii-reagents ezinetyhefu kakhulu.Iya kubangela umonakalo ongenakulungiseka kumzimba womsebenzisi, kwaye ibangele nomonakalo othe ngqo ngexesha lovavanyo.
Kwangaxeshanye, kwabo banenani elikhulu leesampuli zokufunda, ukuhlukana kunye nokutsalwa kwe-nucleic acid ngumsebenzi onzima kakhulu.
Ukwahlukaniswa kwe-Nucleic acid kunye neekhithi zokutsalwa kwimarike ngoku zivuthiwe kwaye kukho iibrendi ezininzi, kodwa ziyafana.Nokuba yi-silica gel membrane column centrifugal kit okanye i-magnetic bead method kit, ithatha ixesha elininzi kwaye iyabiza.Ukongeza kwiindleko zekiti, kukho kwakhona iimfuno ezikhethekileyo zezixhobo zebhubhoratri.Indawo yokusebenza ezenzekelayo esetyenziswa kwindlela yemagnethi yentsimbi sisixhobo esiqhelekileyo somgangatho ophezulu wexabiso eliphezulu, eyindleko enkulu kwilabhoratri.
Isishwankathelo
Ngaphambi kokuqhuba iimvavanyo ze-PCR, ukunyangwa kwangaphambili kweesampuli yinto engenakuphepheka kwaye ihlala iyintloko kubaphandi.Indlela yokusombulula le ngxaki kunye nokuba iimvavanyo ze-PCR zingenziwa ngaphandle kokwahlulwa kunye nokutsalwa kwe-nucleic acids ibisoloko icinga uninzi lwabaphandi besayensi kunye nabasebenzi belabhoratri yeklinikhi.
Isisombululo seForegene
Emva kweminyaka yophando olunzima kwithekhnoloji ye-PCR ethe ngqo kunye nezixhobo ezinxulumeneyo, i-Forgene yaqhawuka ngempumelelo kwiibhotile ezininzi kwaye iphumelele ngokuthe ngqo kwi-PCR kwiintlobo ezininzi zeesampulu ezahlukeneyo ezinokuxhathisa okunamandla kunye nokuguquguquka, ukuvumela abaphandi ukuba balahle ukwahlukana okunzima kunye nobungozi kunye nokutsalwa kwe-nucleic acid.Oku kuya kunciphisa kakhulu amandla okusebenza komntu wonke, kukhawulezise inkqubo yovavanyo, kwaye konge uphando lwezenzululwazi kunye neendleko zovavanyo.
Ukuqonda kukaForgene kunye nolwazi lweDirectPCR
Okokuqala, itekhnoloji ye-DirectPCR yitekhnoloji ye-PCR ethe ngqo yeesampulu ezahlukeneyo zebhayoloji.Ngaphantsi kwesi simo sobugcisa, akukho mfuneko yokwahlula kunye nokukhupha i-nucleic acids, kwaye isampuli ye-tissue isetyenziswe ngokuthe ngqo njengento, kwaye i-primer gene ejoliswe kuyo yongezwa kwi-PCR reaction.
Okwesibini, itekhnoloji yeDirectPCR ayisiyotekhnoloji yokukhulisa itemplate yemveli yeDNA kuphela, kodwa ikwabandakanya iRNA template reverse transcription PCR.
Okwesithathu, itekhnoloji ye-DirectPCR ayenzi ngokuthe ngqo kuphela ukusabela okusemgangathweni kwe-PCR kwiisampulu zethishu, kodwa ikwabandakanya ukuphendula kweXesha lokwenyani le-qPCR, efuna inkqubo yokusabela ibe namandla okumelana nokuphazamiseka kwe-fluorescence yangasemva kunye nokuchasana nezicimi ze-endogenous fluorescence.
Okwesine, iisampulu zezicubu ezijoliswe kwi-teknoloji ye-DirectPCR zifuna kuphela ukukhululwa kwee-templates ze-nucleic acid kwaye zingasusi iiprotheni, i-polysaccharides, i-ion yetyuwa, njl.Oku kufuna i-nucleic acid polymerase kunye ne-PCR Mix kwinkqubo yokusabela ukuba ibe ne-anti-reversible egqwesileyo kunye nokuguquguquka, kwaye inokuqinisekisa umsebenzi we-enzyme kunye nokuphindaphinda ukuchaneka phantsi kweemeko ezinzima.
Okwesihlanu, iisampulu zezicubu ezijoliswe kwithekhnoloji ye-DirectPCR azizange zithotyelwe naluphi na unyango lokutyebisa i-nucleic acid, kwaye ubungakanani betemplate buncinci kakhulu, obufuna uvakalelo oluphezulu kakhulu kunye nokukhulisa ukusebenza kakuhle kwenkqubo yokusabela.
Ukuqukumbela
I-teknoloji ye-DirectPCR yenye yezona zinto zibalulekileyo zophuhliso lwezobuchwepheshe kunye nezinto ezintsha kwiminyaka eyi-30 edlulileyo ukususela ekuzalweni kobuchwepheshe be-PCR.UForgene une kwaye uya kuqhubeka nokuba nguvulindlela kunye nomqalisi wobu buchwepheshe.
Ithemba lokusetyenziswa kweteknoloji yeDirectPCR ibanzi kakhulu.Ukuphuculwa okuqhubekayo kunye nokukhuthazwa kobu buchwepheshe ngokuqinisekileyo kuya kuzisa utshintsho oluphazamisayo kuphando lwezenzululwazi kunye nomsebenzi wokuhlola.Olu lutshintsho lweteknoloji yePCR.
Ixesha lokuposa: Feb-21-2017
