1. Hydrolyse the peptide refluxing with 6 moldm-3 hydrochloric acid for 24 hours 2. Wearing gloves, use a ruler to draw a pencil line 1.5cm from the bottom of the plate 3. Use a capillary tube to apply a small drop of the solution of amino acids to the mid-point of the pencil line 4. Leave drops to dry completely 5. Add solvent to a beaker at a depth of no more than 1cm 6. Place TLC plate in the beaker 7. Place a lid on the beaker (to keep inside saturated with solvent vapour) 8. Allow solvent to rise up the plate until the solvent reaches near the top of the paper 9. Remove the plate and immediately mark the position of the solvent front with a pencil 10. Let the plate dry 11. Locate the amino acid spots using a locating agent/ UV 12. Measure the Rf values of each amino acid & compare these to known values
