Experiments

Experiment Overview

Here we will outline in words the basic progression of each of our experiment in order of experiments conducted. If specific perimeters were used in each experiment note them.

Preparation of Competent Cells

Here include information on both the DH5a and BL21 competent cell prep.

pET-28a isolation

Here include information how we isolated and purrified pET-28a from DH5a/pET-28a

Site Directed Mutagenesis of pET-28a

Here include information on how we mutated pET-28a to insert PstI RE site.

Confirmation of SDM Success

Here include information on the steps we took to confirm that our SDM occurred correctly.

Isolation of pET-28a(PstI) from DH5a

Here include information on how we isolated and purrified pET-28a(PstI) from DH5a/pET-28a

To be, or not to be: that is the question: Whether ’tis nobler in the mind to suffer The slings and arrows of outrageous fortune, Or to take arms against a sea of troubles, And by opposing end them? To die: to sleep; No more; and by a sleep to say we end The heart-ache and the thousand natural shocks That flesh is heir to, ’tis a consummation Devoutly to be wish’d. To die, to sleep; To sleep: perchance to dream: ay, there’s the rub; For in that sleep of death what dreams may come When we have shuffled off this mortal coil, Must give us pause: there’s the respect That makes calamity of so long life; For who would bear the whips and scorns of time, The oppressor’s wrong, the proud man’s contumely, The pangs of despised love, the law’s delay, The insolence of office and the spurns That patient merit of the unworthy takes, When he himself might his quietus make With a bare bodkin? who would fardels bear, To grunt and sweat under a weary life, But that the dread of something after death, The undiscover’d country from whose bourn No traveller returns, puzzles the will And makes us rather bear those ills we have Than fly to others that we know not of? Thus conscience does make cowards of us all; And thus the native hue of resolution Is sicklied o’er with the pale cast of thought, And enterprises of great pith and moment With this regard their currents turn awry, And lose the name of action.–Soft you now! The fair Ophelia! Nymph, in thy orisons Be all my sins remember’d.

Protocols

Here we will include a detailed list of all the basic protocols used in our experiments. Specific parameters for things such as PCRs will be noted above

To be, or not to be: that is the question: Whether ’tis nobler in the mind to suffer The slings and arrows of outrageous fortune, Or to take arms against a sea of troubles, And by opposing end them? To die: to sleep; No more; and by a sleep to say we end The heart-ache and the thousand natural shocks That flesh is heir to, ’tis a consummation Devoutly to be wish’d. To die, to sleep; To sleep: perchance to dream: ay, there’s the rub; For in that sleep of death what dreams may come When we have shuffled off this mortal coil, Must give us pause: there’s the respect That makes calamity of so long life; For who would bear the whips and scorns of time, The oppressor’s wrong, the proud man’s contumely, The pangs of despised love, the law’s delay, The insolence of office and the spurns That patient merit of the unworthy takes, When he himself might his quietus make With a bare bodkin? who would fardels bear, To grunt and sweat under a weary life, But that the dread of something after death, The undiscover’d country from whose bourn No traveller returns, puzzles the will And makes us rather bear those ills we have Than fly to others that we know not of? Thus conscience does make cowards of us all; And thus the native hue of resolution Is sicklied o’er with the pale cast of thought, And enterprises of great pith and moment With this regard their currents turn awry, And lose the name of action.–Soft you now! The fair Ophelia! Nymph, in thy orisons Be all my sins remember’d.

Refrences

Rose, D. This is a test (2017). Sci. Awesome. 28-29

Team:U of Guelph/Experiments