Question

i nee help analysing this data

Anssume that the fractions you collected from the partial purification by ionic chromatography were incubate with lambda DNA and EcoR1 reaction buffer( Restriction enzyme reactions) and you obtained the following bands in your agarose gel.

2 3 4 5 6 Lane Fraction Salt Content B and Lamda EcoR1 Marker 2 None 3 Lamda + fraction 2 Lamda + fraction 3 No salt No salt 21 3 4 5 7 8 9 10 6 6 7 8 9 7 8 Lamda + fraction 5 Lamda + fraction 6 Lamda + fraction 7 Lamda + fraction 8 Lamda + fraction 9 0.1M KCI 0.2M KCI 0.2M KCI 0.5M KCI 0.5M KCI IL 10 9

Answer 1

1. Once the DNA have been separated it can be viewed by placing the gel under UV light in a Gel Doc machine, due to the presence of dye the bands of DNA fragments glow.

2. Fraction6 and 7 have EcoR1 fragments after partial purification as most of the lighter bands generated have same size as that of the marker. The peak activity of EcoR1 could be seen in fraction 6 as maximum fragments are generated.

3. Analysing the data it can be interpreted that fractions1, 2 and 3 have similar sized fragments of DNA, therefore action of the enzyme in the presence of 0.1M salt and no salt are similar. For fraction 4, the fragments are seen to overlap due to similar size, the same is in case of fraction 8 and 9, and the separated fragments are seen as overlapping bands. In case of fraction 6 and 7, the fragment size is similar and maximum similarity of the fragment size can be found when compared to the marker.