Chemical Biology 3OA3, September 2009 Page 45 i.Characterization of product peptide:TLC:In this experiment, TLC will be used to check the purity of the peptide. Fine capillaries for applying spots will be provided, or else can be made from Pasteur pipettes. Your T.A. will demonstrate the technique. You can also review the CHEM 20A3 Lab Manual at this point. 1.Place 1-2 mL of the developing solvent (chloroform : methanol : acetic acid (30:15:5 v/v)) in the chromatography jar, cover tightly and allow to stand 5-10 minutes before using. 2.Remove a small sample of your peptide (enough to see) and dissolve in 2-3 drops of methanol in a vial. 3.Make up a similar solution of the Gly-Ala peptide standard provided. 4.Use the silica gel TLC sheets provided. With a pencil mark two dots about 6 mm apart, and 8-10 mm from the bottom of the plate. On the first mark, place a spot of your control peptide solution. On the second mark, place a spot of your product solution. When each plate has been spotted, place the plate, spotted end down, in the developing jar.
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