Which one of the following attributes of the green fluorescent protein (GFP) make it a suitable candidate for hydrophobic interaction chromatography (HIC)?

Before you can add your recombinantly expressed GFP to the HIC column for purification, you needed to lyse the cells.  Which one of the following statements best explains why you did this step?

You are the head of a biotechnology lab that has a number of therapeutic proteins that could be used to treat COVID-19. There are a number of candidate proteins that you could potentially make. These proteins are listed below. Given the information about each protein, choose what you think would be the optimal system to express the protein.

When transforming bacteria with a plasmid, we are often interested in how many plasmids have (on average) ended up in each bacterial cell (i.e. the copy number).

We can use quantitative PCR to do this, where we are interested in the cycle number when the amplified product crosses a predetermined threshold (Cq) as shown below:

This will enable us to compare the threshold (Cq) values for genes on the plasmid with genes on the bacterial chromosome.

In this scenario, you have transformed E. coli with the plasmid pCOOKIE.  The plasmid carries the Amp gene, while the E. coli contains the SAM gene on its chromosomal DNA.  You have primers for both the Amp and the SAM genes and you use these to perform quantitative PCR on your transformed cells.

You amplify the Amp and the SAM genes in triplicate using quantitative PCR and get the following results:

The equation for calculating copy number of a plasmid is:

Using the data provided, what is the copy number for pCOOKIE in your experiment?

* Round your answer to the nearest whole number; figures only; no units.

When performing laboratory experiments, we often find that our actual results are not quite the same as the ideal or expected results.  Look at the following results from the qPCR assay (these students used the same protocol that you used).  Which one gives the most reliable results?

In the previous question you were asked to calculate the transformation efficiency from the equation given.  Which one of the following options would be the correct units for that calculation?

You have transformed some E. coli  with a plasmid vector (pCOOKIE) containing an ampicillin resistance gene.

You perform the transformation as follows:

1. 10 µL of a 10 µg/mL stock of plasmid is added to the transformation tube.
2. The total volume in this tube is 100 µL.
3. All 100 µL from the transformation tube is spread onto the agar plate.

The plates are incubated and the results are shown in the table below:

Using the results shown in the table and the transformation efficiency formula below, calculate the transformation efficiency for pCOOKIE in these cells.

Give your answer as the nearest whole number (no decimal points), figures only, no units.

For the following restriction enzymes (REs), the recognition sequences and the cut sites are shown on the template strand (all reading left to right as 5' to 3'), indicate which REs create sticky ends and which ones create blunt ends.

Cytosine makes up 20% of the nucleotides in a particular mammalian gene.  What percent of the nucleotides in this gene will be thymine?

Your objective in a cloning experiment is to move a gene of interest between various plasmid vectors to optimise various parameters at different stages, such as propagation of the plasmid vector and protein expression.  Which one of the following cloning technologies would be most suitable for this purpose?