Question

THIS IS A CONCEPTUAL UNDERSTANDING QUESTION: Grapes have a lot of DNA in them. However, when you used the same protocol, you did not obtain any DNA from the grapes. What can you do differently so that you could get DNA from your grapes?

Answer 1

Answer:

As per the DNA isolation process, many steps available their many chances are present those create mistakes:

a. Sample crushing in liquid nitrogen: it’s a first step in which sample take in a motor pestle and crush solely and single side rotation smoothly otherwise cell will rapture and it makes DNA damage.

b. Buffer adding and maintain temperature and time: after making sample in powder form add buffer add in equal quantity.

c. Organic components mixing and separating sugar and proteins contents: at this step PCI (Phenol Chloroform Isoamyl alcohol) and CI (Chloroform Isoamyl alcohol) add for sugar and protein removing. In this step, two-layer make and we need to take the upper layer for further process. In some time, it’s a great mistake for DNA lose.

d. Maintaining temperature and washing step: Mostly sodium acetate and amyl alcohol use for overnight treatment but due to shortage time, some person does this process continuously that make mistake. Washing process also does by 70%, 80%, 90% and ten 100% alcohol in this manner otherwise DNA can damage.

e. Dissolving step: in this procedure always use double distilled water (DDW) but at this step, we should use fresh and DDW to avoid contamination.

f. DNA concentration check by electrophoresis or nano-drop: DNA concentration checking by nanodrop have a low risk for DNA damage but this is a costly technique and easily not available each place. Mostly for DNA check, electrophoresis utilizes. Hence electrophoresis little change buffer pH will create big damage.

Conclusion:

At last, we can say, DNA isolation not an easy method. Each step is a crucial step. Therefore, to isolate fine quality DNA, take all step seriously and with concentrate mind.