Bryan Hernandez: Difference between revisions

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For information on me, see [http://thebryanhernandezgame.wordpress.com/ The Bryan Hernandez Game].
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== Bio ==
== Bio ==


*I am currently an undergraduate researcher in the [[Keasling Lab]] in Lawrence Berkeley National Laboratory.  My project is [[IGEM:UC Berkeley/2006| iGEM]] 2006.
*I am currently an undergraduate researcher in the [[Endy Lab]] at MIT.  I am currently building and characterizing orthogonal riboregulators.  Learn more about riboregulators and the work I have done [http://parts2.mit.edu/wiki/index.php/Berkeley2006-RiboregulatorsMain here]
*MIT Class of 2009; Majoring in Mathematics and Biological Engineering.
*MIT Class of 2009; Majoring in [http://math.mit.edu/| Mathematics] and [http://web.mit.edu/be/index.htm| Biological Engineering].


== Projects ==
== Projects ==
===[[IGEM:UC Berkeley/2006 | iGEM UC Berkeley]]===


Our project is to create an addressable cell-to-cell communication mechanism in e. coli.<br>
===Construction and Characterization of Riboregulators===
[[IGEM:UC Berkeley/2006/bryans notebook|notebook]]<br>
A Riboregulator is a post-transcriptional regulator used in bacteria.  Its mechanism for regulation involves the occlusion of the Ribosome via a hairpin on the mRNA transcript in the 'off' state thereby preventing (or 'locking') translation of the ORF.  The 'on' state is recovered by introducing a complementary sequence (or 'key') that disrupts the hairpin on the mRNA transcript allowing the Ribosome to bind and initiate translation.  To learn more, go [http://parts2.mit.edu/wiki/index.php/Berkeley2006-RiboregulatorsMain here].
[[IGEM:UC Berkeley/2006/bryans -80 stocks|-80 stocks]]
 
*[[Bryan Hernandez/UROP Proposal|UROP Proposal]]
 
===[http://parts2.mit.edu/wiki/index.php/University_of_California_Berkeley_2006 Addressable Conjugation in Bacterial Networks]===
*Our project was to create an addressable cell-to-cell communication mechanism in e. coli.<br>
*[[IGEM:UC Berkeley/2006 | iGEM UC Berkeley]]
*[[IGEM:UC Berkeley/2006/bryans notebook|notebook]]<br>


===[[Sortostat]]===
===[[Sortostat]]===
The Sortostat is a microfluidic chemostat integrated with a cell sorter.  My project consists of demonstrating the sortostat's chemostat functionality, a technique rarely seen in microfluidics. Chemostasis must first be demonstrated before any meaningful demonstration of the sortostat's sorting capabilities can be shown. The sortostat's sorting ability is limited by the extent to which it can make optical descriminations between cells.  Therefore, in order to demonstrate its sorting ability, two visually different populations of cells will be grown to steady state after which sorting is initiated.  If successful the populations will clearly diverge in number, the one sorted against will diminish well below its steady state level and the one preserved should rise by the same amount that the other falls. The total population of cells in the chemostat should be conserved in the end if chemostasis was maintained throughout sorting.          
The Sortostat is a microfluidic chemostat integrated with a cell sorter.  My project consists of demonstrating the sortostat's chemostat functionality, a technique rarely seen in microfluidics.  
 
'''Immediate Goals'''
*Debug all engineering-related problems with sortostat.
*Demonstrate Sortostat's ability to attain chemostasis in a population of E. Coli.
*Demonstrate Sortostat's ability to sort two phenotypically different populations of E. Coli.  In this case, I will be using a mixture of E. Coli expressing CFP and YFP.  
*Demostrate Sortostat's ability to select for an arbitrary population of cells that are phenotypically different from others and maintain this population at a steady state.
*[[Bryan Hernandez/UROP Proposal|UROP Proposal]]
*[http://www.midgard.liu.se/~b00perst/chemostat.pdf Chemostat Theory]
*[http://www.midgard.liu.se/~b00perst/chemostat.pdf Chemostat Theory]
*[[Sortostat/Experiments]]
*[[Sortostat/Experiments]]
*[[Sortostat/Growth Tests]]
*[[Sortostat/Growth Tests]]
...


'''Project Goals'''
-----------
 
*[[Bryan Hernandez/UROP Proposal|UROP Proposal]]


*[[Sortostat]]
[[Bryan Hernandez/20.109|20.109]]<br>
[[IGEM:UC Berkeley/2006/bryans -80 stocks|-80 stocks]]<br>
[[media:oligos.xls|oligos]]


#Debug a microfluidic chemostat ([[Sortostat]]) to improve the time-varying specific selection of cell populations.
contact me at bryanh (AT) mit
#*Currently troubleshooting problems:  i) Cell Death after 3-4 days. This problem has not been an issue for the past few runs that lasted over 300 hours.  We initially thought the cell death was due to oxygen depletion however if this was the case we'd expect to see this more consistantly. and ii) Inaccurate cell counts due to poor image processing.  There is still much to be done to resolve this issue.  Perhaps using software such as Cell Profiler will help to achieve more accurate cell counts.
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#Evaluate the response of populations of E.Coli cells containing engineered genetic circuits (http://parts.mit.edu) to particular selective pressures using the Sortostat.

Latest revision as of 07:48, 18 May 2010

For information on me, see The Bryan Hernandez Game.

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