Biochem Report

Pre-lab

1. One of the factor that effect the movement of molecules in paper electrophoresis would be the molecule itself where the size, shape, charge and chemical composition of the molecules to be separated will affect the movement (Boyer, 2006). Besides that, the movement of molecules is the chemical composition of the buffer (Deyl, Everaerts and Prusík, 1979). Other factor that could affect the movement of molecules are endosmotic flow and wick flow (Biswajit and Basu, 2006). Applied voltage when running the electrophoresis might affect the movement of the molecules too. The higher the voltage applied, the faster the movement of molecules.

1. Paper electrophoresis is not suitable to use to separate protein because proteins are considered macromolecules. Besides, the absorption and surface tension associated with paper electrophoresis usually alter or denature the macromolecules, thus it will cause a poor resolution (Switzer and Garrity, 1999).

1. Table 1: Shows the net charge of lysine, histidine and aspartic acid in pH 7.6 and pH 3

1. Lysine

Histidine

Aspartic acid

1. Number of moles of aspartate   = 10 x 10-3 x 1L

                                      = 0.01 moles

5 µmoles = 5 x 10-6 moles

Volume of sample in 5 µmoles of aspartate = [pic 1]

                                                  = 5x 10-4 L

                                                  = 0.5 mL

Result [pic 2]

Figure 1: Shows the image of the bands of the DNA ladder and sample which is HindIII digest of that λ phage DNA that appeared in the agarose gel under UV light

[pic 3]

Figure 2: Shows size of DNA ladder used in experiment

Table 2: Shows the fragment size of DNA ladder (kbp), distance travelled by DNA ladder(cm) and the log10 (fragment size) of DNA ladder.

[pic 4]

Figure 3: Graph of distance travelled by the bands (cm) against log10 (fragment size) of DNA