Solid Phase peptide synthesis(SPPS) PLC

Fmoc-resin
Polystyrene AM rRAM 0.62 mmol/g

pyperidine 

Treat Fmoc-resin with 20% piperidine() in DMF 2 times 3mL/100mg. use 160 mg Fmoc resin so used 4,8mLto wash. Wash the resin 5 time DMF 3mL/100mg resin each time. Use distilled DMF in the final wash.
DMF Dimethylformamide: common solvent for chemical reaction.
1st step: Fmoc removal

------>>
 Fmoc-gly-OH

MW = 297.3

Fmoc-Leu-OH 

 MW = 353.4
Fmoc-Pro-OH 

MW = 337.37

HOBt-Hydroxybenzotriazole

DIC MW= 126.20

Step 2 Coupling
 4 equivalens(0.4 mmol X 297.4 = 0.119 g) Fmoc-gly-OH + 6(0.6 mmol x 337.37=0.202 g ) equivalents HOBt() in distilled DMF(1ml/100mg resin)= 1.6 mL. add 4 equivalents (0.4 mmol x 126.20 = 0.050 g)DIC, vortex briefly and and the mixture to the resin.
Add 0.0005 equivalents Bromophenol blue(50 uL) and start vortex shaking.---the reactions turns from blue to yellow. Filter of the reaction mixture and wash the resin 5 times with DMF(3ml-100mg resin each time).
Fmoc-gly-Resin
-then Fmoc removal again then coupling step with  Fmoc-Leu-OH
- Fmoc remoal then coupling step with Fmoc-Pro-OH , 
PLG-Resin
The final step wil deprotecting and cleavage the solid phase.
Deprotection and cleavage from the solid phase:
Wash the resin 5 times with DMF (3mL/100 mg resin)
- wast the resin 10 times with CH2Cl2 (3mL/100 mg resin)
- evaporate the solvent, add cleavage mixture(TFA:H2O:Thioanisol:ethanedithiol=8.75:0.5:0.5;0.25) to the resin (2mL/100 mg resin), let the mixture stand without stirring for 4 h.
- filtrate the mixture and wash the resin with TFA(3mL/100 mg resin, 4,8mL) and rotary evaporation. 
- Evaporate the mixture with 3mL HOAc 2 times. collect the crystalline. optional steps: triturate 2 or more the peptide by adding ice-cold 10mL Et2O, after standing 5 min withdraw the ether.
Yield: 30 mg PLC