User:JeffreyLau: Difference between revisions

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== About ==
== About ==
Undergraduate concentrating in Computer Science (class of 2007), member of 2006 Harvard iGem team.
Undergraduate concentrating in Computer Science (class of 2007), member of the [[IGEM:Harvard/2006|2006 Harvard iGem team]]. I'm from Sunnyvale, CA.


== 2006/06/15 ==
== Contact ==
'''Ligation'''
*Since our e-gel failed, we ligated DNA extract from David and Peng.
*Mixed 6 µL insert (E0241), 2 µL vector (R0010), 2 µL DNA dilution buffer in reaction vial
*Added 10 µL ligation buffer
*Added 1 µL ligase, mixed
*Let incubate 5min at room temperature


'''Transformation'''
jlau at fas dot harvard dot edu<br>
*Mixed 20 µL of ligation product with 30 µL competent cells
discobird at gmail dot com
*Negative control: 2 µL of dH20 with 30 µL competent cells
*Positive control: 5 µL of positive control with 30 µL competent cells
*Incubate on ice for 20min
*Heat shock @42C for 30s
*Rest on ice for 2min
*Incubate overnight @37C


== 2006/06/14 ==
== Projects ==


'''Biobrick tutorial'''
I am working on the [[IGEM:Harvard/2006/Cyanobacteria|cyanobacteria oscillator project]].


''More detailed procedure notes are in lab notebook.''
== Lab notebook ==
<calendar>
name=User:JeffreyLau/Notebook
date=2006/07/01
view=threemonths
format=%name/%year-%month-%day
weekstart=14
</calendar>


*Began with 12 samples (3x each of negative control, R0010, E0241, E7104). Fortunately our negative controls showed no growth overnight.
=== Quick links to notebook entries ===
[[User:JeffreyLau/Notebook/2006-6-14|2006-6-14]]: Notes on using the nanodrop


*Made 9 minipreps for the remaining samples. Set aside the E7104 samples (they are a positive control).
[[User:JeffreyLau/Notebook/2006-6-15|2006-6-15]]: Ligation protocol, transformation protocol


*Performed digests on 2x each of R0010 and E0241. We lost one of our E0241 samples by accident, so we threw out an R0010 sample to balance.
[[User:JeffreyLau/Notebook/2006-6-20|2006-6-20]]: Digest protocol
 
*Added Calf Intestinal Phosphatase (CIP) to R0010 samples to prevent self-ligation of the R0010 vector.
 
*Performed gel electrophoresis on R0010 and E0241 samples.
**Our e-gel was not entirely successful.
[[Image:HH-JL biobrick-tutorial egel.jpg]]
 
Must be more careful next time.
 
'''Notes on using the nanodrop'''
*Nanodrop measures concentration of DNA in small samples (1 µL). It is apparently not very accurate.
*Clean nanodrop thoroughly with kimwipes and ethanol, before and after each use
*On desktop, "ND-1000"->"Nucleic Acid"
*Before each set of samples:
**Calibrate nanodrop with drop of distilled water
**Calibrate nanodrop with drop of background (solute), e.g. distilled water or elution buffer
*Add sample to nanodrop, enter name of sample, "Measure"
**Note: mix sample before placing on nanodrop

Latest revision as of 10:37, 27 June 2006

About

Undergraduate concentrating in Computer Science (class of 2007), member of the 2006 Harvard iGem team. I'm from Sunnyvale, CA.

Contact

jlau at fas dot harvard dot edu
discobird at gmail dot com

Projects

I am working on the cyanobacteria oscillator project.

Lab notebook

<calendar> name=User:JeffreyLau/Notebook date=2006/07/01 view=threemonths format=%name/%year-%month-%day weekstart=14 </calendar>

Quick links to notebook entries

2006-6-14: Notes on using the nanodrop

2006-6-15: Ligation protocol, transformation protocol

2006-6-20: Digest protocol

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