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Isisetshenziswa sokuhlanza se-Oligo se-Oligo primer synthesis Isithombe esifakiwe
  • Isisetshenziswa sokuhlanza i-Oligo se-Oligo primer synthesis

Isisetshenziswa sokuhlanza i-Oligo se-Oligo primer synthesis

Isicelo:

Imishini yokuhlanza uketshezi ezenzakalelayo ivumela ukudluliswa kwenani loketshezi oluhlukene.Uketshezi lushaywa noma lushiswe ngokuhlanganiswa noma amakholomu okuhlanza e-C18.Idizayini edidiyelwe, isistimu yokulawula i-axis eyodwa kanye nesixhumi esibonakalayo esikahle somshini womuntu sinika amandla okulawula okuzenzakalelayo ngokuphelele kwemishini.


Imininingwane Yomkhiqizo

Omaka bomkhiqizo

Isisetshenziswa sokuhlanza i-Oligo se-Oligo primer synthesis,
umshini wokusekela we-oligo synthesis,

Ukucaciswa

Amabhodi ahambisanayo 1, 3, 5, 8.
Ukuhlunga ukuhlungwa kwe-blow, ukuhlunga kokudonsa
Inombolo yezimbobo zokujova 5, 6, 7, 8, 9, 10.
Izinhlobo zamapuleti ahambisanayo Ipuleti le-C18, ipuleti elijulile, ipuleti lokwenziwa (lihambisana namapuleti amaningi okwenziwa), ipuleti le-microtiter
Imojuli i-eksisi eyodwa noma i-eksisi ekabili
Amandla kagesi 220V
Iwaranti 1 unyaka
Ngokwezifiso Kwamukelwe

Izinsiza Zokuhlanzwa zezindaba zokuhlanza i-Oligo3

Izindlela ezahlukene zokuhlanza

1. Gel Electrophoresis Chromatography Ukuhlanzwa
Sebenzisa i-denaturing polyacrylamide gel electrophoresis chromatography ukuze uhlanzeke.I-ejenti ye-denaturing ngokuvamile i-4M formamide noma i-7M urea, ukugxila kwe-acrylamide kuphakathi kuka-5-15%, futhi ingxenye ye-methacrylamide iphakathi kuka-2-10%.
Ngemuva kwe-electrophoresis, indawo yebhande le-nucleic acid idinga ukunqunywa ngaphansi kwemisebe yokukhanya kwe-ultraviolet, ijeli equkethe i-nucleic acid ehlosiwe iyanqanyulwa, i-nucleic acid iphihlizwe futhi ifakwe, bese isixazululo se-leaching sigxiliwe, sikhishwe usawoti, quantified futhi lyophilized.

2. I-DMT-On, i-HPLC Purification
Khetha imodi ye-DMT-On ngesikhathi sokuhlanganiswa, umkhiqizo ongahluziwe wawufakwe phakathi futhi ugxiliswe ekamelweni lokushisa ukuze kukhishwe i-ammonia eningi ngemva kwe-aminolysis.
Ukuhlukaniswa kwenziwa kusetshenziswa ikholomu ye-C18 ene-acetonitrile kanye ne-10% ye-triethylamine-acetic acid (TEAA) njenge-elient.Ngemva kokuqedwa kwe-lution, kuyagxiliswa, bese iqembu le-DMT lisuswa nge-trifluoroacetic acid.Ngemuva kwe-neutralization, amanye ama-salts nama-molecule amancane akhishwa ngeshubhu elinqunyiwe, futhi ekugcineni akhishwe usawoti.

Le ndlela ingathola umkhiqizo ngokuhlanzeka okuphezulu, kodwa kuyadingeka ukuba unake ukwenzeka kwe-depurination.

3. I-DMT-Off, i-HPLC Purification
Khetha i-DMT-Off ngesikhathi sokuhlanganiswa, futhi umkhiqizo ongahluziwe wawugxiliswe phakathi futhi ugxiliswe ekamelweni lokushisa ukuze kukhishwe i-ammonia eyengeziwe ngemva kwe-ammonolysis.
Ukwehlukaniswa kwenziwa kusetshenziswa ikholomu ye-C18 ene-acetonitrile kanye ne-10% ye-triethylamine-acetic acid emanzini njengokulula.Ngemuva kokuthi ukuhlukaniswa kuqediwe futhi kunqunywe, ama-aliquots a-lyophilized.

Le ndlela idinga ukulungiswa ngokucophelela kwezimo zokuhlukaniswa, futhi ingathola nama-molecule ahlosiwe amsulwa.

Imishini ye-oligo synthesis accerry yomjikelezo we-dna rna synthesis, evumelana ne-primer evamile, nezinye i-primer chain primer, yenza i-oligo ibe msulwa kakhulu.
umkhiqizi wase-china wemishini ye-oligo synthesis enekhwalithi enhle kanye nenkonzo engcono kakhulu.

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