a from the photograph of your gel estimate the size and hoge

a. from the photograph of your gel, estimate the size and hogeneity/purity of your amplified DNA samples?

b. how do you match the bands in the standard to their molecular weight?

c. do you think your PCR captured a single sequence?

d. Make a graph of log MWv. distance traveled by the DNA standards.

(1st well is Indicators, 2nd well PCR, 3rd well - undigested, 4th well - digested)

Solution

Ans a. The size of one band is beyond 10 kb where as the lower band is of 5 kb in size as compared to the standard. With respect to the homogeneity is concerned the DNA is pure and free from contamination of proteins as there are no remains near the well or glowing inside the well. This shows the homogeneity of the DNA.

Ans b. The PCR amplified bands in the well 2 is compared with that of the standard ladder (Each band will be labeled starting for the lowest band (away from the well) to the highest (near to the well)). The position of the band in the 2 well is compared with the standard molecular weight. For example in the second well it shows from the gel the first band near to the well is higher than10kb fragment, where as the second band is 10kb as it is same distance from well compared to the standard ladder. It is measured as the distance travelled from the well which shows the molecular weight and the number of Nucleotides present in each band.

Ans c. No. Even though a same sequence has increased exponentially of different length at different places, it has captured variable length sequences. Hence it has produced two different sizes of the band.