Reference no: EM133357278
Questions:
1. There are three major steps in PCR process, denaturation, annealing, and extension. In the lab, you will run the denaturation step at 94°C, and you should get the PCR product as desired. Let's say you accidentally make a mistake and setup the PCR machine to do the denaturation step at 84°C. You find out you get no product. Why does this make sense? (e.g. what is the likely reason for the lack of a PCR product?)
2.Alternatively, let's say you accidently set the annealing temperature in the PCR machine to 75°C, rather than the 65°C given in the manual. What problems might this create and why?
3) What is the name of for the specific sequence of E. coli that will be amplified in this experiment, and why does it make the detection specific to E. coli (e.g. why wouldn't it give a band if you had B. subtilis contamination)?
4. You have a sample with two sizes of DNA fragments, 1000 base pairs (bp) and 5000 bp. You run the DNA agarose gel and get the picture below. Which band corresponds to the 1000 bp and which the 5000 bp fragment and why?
1. What would happen if the column is dried during the Isolation and Determination of mCherry Protein by Using Affinity Chromatography
2. What is polyhistidine-tag (His-tag)? How can a protein containing His-tag be purified by using affinity chromatography?
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