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a 1 base 5'-OH and a 3'-OH of the next base form a phosphite triester,
3. Remove the protective agent on the base with concentrated ammonium hydroxide under heating
Introduction to the principle of synthesis of DNA synthesizer
Synthetic principle of DNA synthesizer:
Solid phase synthesis of DNA: the 3' end of the DNA is immobilized on a substrate, and then nucleotides are sequentially added in the 3' to 5' direction until the desired DNA fragment is synthesized. It is different from DNA synthesis using DNA polymerase.
Synthesis:
The 3' end of the first base is immobilized on the resin, the 5'-OH of the next base is protected with di-p-trimethoxymethyl DMT, the amino group at the base is protected with benzoic acid, and then 3'- OH is activated with an aminophosphoric acid compound.
2. Then oxidize to form a phosphate triester with iodine
3. Add dichloroacetic acid to remove the protective agent DMT on the second base 5'-OH
Cycling after adding the next base synthesis
1. Removal of the protective agent DMT on 5'-OH with thiophenol
2. Disconnect the fragment from the solid resin with concentrated ammonium hydroxide and elute
4. Remove ammonium hydroxide, vacuum dry
5. Liquid chromatography or PAGE, the longest recovery fragment.
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