Revision as of 15:08, 25 October 2017 (view source) Drose (Talk | contribs) ← Older edit		Revision as of 17:23, 25 October 2017 (view source) Drose (Talk | contribs) Newer edit →
Line 90:		Line 90:
	<h1 class="descSub">Project Overview</h1>		<h1 class="descSub">Project Overview</h1>
	<p class="descP">		<p class="descP">
−	Here we will write our over arching plans for our project without going into too much detail at every step.<br><br>	+	Step 1: Cloning of <i>frc</i> and <i>oxc</i> into DH5&alpha;
−		+	<ul>
−	To be, or not to be: that is the question:	+	<li>Synthesise <i>frc</i> and <i>oxc</i> </li>
−	Whether ’tis nobler in the mind to suffer	+	<li>Add PstI cut site to pET-28a </li>
−	The slings and arrows of outrageous fortune,	+	<li>Ligate <i>frc</i> and <i>oxc</i> into pET-28a </li>
−	Or to take arms against a sea of troubles,	+	<li>Transform pET-28a<i>frc</i>/<i>oxc</i> into DH5&alpha; </li>
−	And by opposing end them? To die: to sleep;	+	</ul>
−	No more; and by a sleep to say we end	+	Step 2: Clone <i>frc</i> and <i>oxc</i> into BL21
−	The heart-ache and the thousand natural shocks	+	<ul>
−	That flesh is heir to, ’tis a consummation	+	<li>Purrify pET-28a<i>frc</i>/<i>oxc</i> from DH5&alpha; </li>
−	Devoutly to be wish’d. To die, to sleep;	+	<li>Transform pET-28a<i>frc</i>/<i>oxc</i> into BL21 </li>
−	To sleep: perchance to dream: ay, there’s the rub;	+	</ul>
−	For in that sleep of death what dreams may come	+	Step 3: Express and Purrify FRC and OXC
−	When we have shuffled off this mortal coil,	+	<ul>
−	Must give us pause: there’s the respect	+	<li>Induce expression with IPTG and extract crude proteins</li>
−	That makes calamity of so long life;	+	<li>Purrify proteins using Ni-NTA chromotography </li>
−	For who would bear the whips and scorns of time,	+	</ul>
−	The oppressor’s wrong, the proud man’s contumely,	+	Step 4: Characterize FRC and OXC
−	The pangs of despised love, the law’s delay,	+	<ul>
−	The insolence of office and the spurns	+	<li>Characterize enzyme function using Sodium Oxalate</li>
−	That patient merit of the unworthy takes,	+	<li>Characterize enzyme function using Calcium Oxalate</li>
−	When he himself might his quietus make	+	</ul>
−	With a bare bodkin? who would fardels bear,	+	Step 5: Design a Cleaning Solution and Test on Beerstone
−	To grunt and sweat under a weary life,	+	<ul>
−	But that the dread of something after death,	+	<li>Test ability of enzymes to break down Beerstone</li>
−	The undiscover’d country from whose bourn	+	<li>Design a functional cleaning solution</li>
−	No traveller returns, puzzles the will	+	</ul></p>
−	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.</p>	+
	<h1 class="descSub">The 2017 Aspect</h1>		<h1 class="descSub">The 2017 Aspect</h1>

---

Revision as of 17:23, 25 October 2017

Project Description

Beerstone, FRC and OXC

Calcium oxalate build up (commonly referred to as Beerstone) is a major ongoing problem in the brewing industry. Beerstone is difficult to remove from brewing equipment, requiring the use of harsh cleaning chemicals, and provides a porous, nutrient rich growing environment for bacterial biofilms. Our project is to develop a biological, enzyme based cleaning method for the removal of Beerstone from brewing equipment. We prepose to used the enzymes FRC and OXC from Oxalobacter formigenes oxalate degrading metabolic pathway to breakdown Beerstone.

Project Overview

Step 1: Cloning of frc and oxc into DH5α

• Synthesise frc and oxc
• Add PstI cut site to pET-28a
• Ligate frc and oxc into pET-28a
• Transform pET-28afrc/oxc into DH5α

Step 2: Clone frc and oxc into BL21

• Purrify pET-28afrc/oxc from DH5α
• Transform pET-28afrc/oxc into BL21

Step 3: Express and Purrify FRC and OXC

• Induce expression with IPTG and extract crude proteins
• Purrify proteins using Ni-NTA chromotography

Step 4: Characterize FRC and OXC

• Characterize enzyme function using Sodium Oxalate
• Characterize enzyme function using Calcium Oxalate

Step 5: Design a Cleaning Solution and Test on Beerstone

• Test ability of enzymes to break down Beerstone
• Design a functional cleaning solution

The 2017 Aspect

Here we will write about what we acomplished this year on our project.

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