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The PCR reactions were performed on the Studio Quant5 (ThermoScientific)
More amino acids mean longer synthesis time and higher reagent costs

(7)Fmoc-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin (7) Preparation of Fmoc-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin (6)Fmoc-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin(20DMF)5203DMF3DMF2Fmoc-Ala-OH0.7gDIEA0.4mlTBTU0.56gHOBT0.2gDMF 10ml1.53DMF3DMF2Fmoc-Ala-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin In the Fmoc-Asp(otBu)-Asp(otBu)-Ala-Gly-Leu-Val-WangResin in step (6), add a decapping reagent (20% piperidine in DMF solution), react at room temperature for 5 minutes, and drain, Re-add the capping agent and react at room temperature for 20 minutes, wash with isopropanol for 3 times, DMF for 3 times, re-distilled DMF for 2 times, drain, add Fmoc-Ala-OH0.7g, DIEA0.4ml, TBTU0.56g , HOBT0.2g, DMF 10ml normal temperature reaction 1.5 hours, drained with a vacuum pump, washed 3 times with isopropanol, washed 3 times with DMF, washed 2 times with DMF evaporated, and dried to obtain Fmoc-Ala-Asp(otBu )-Asp(otBu)-Ala-Gly-Leu-Val-WangResin