User:JeffreyLau

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== 2006/06/15 ==
 
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'''Ligation'''
 
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*Since our e-gel failed, we ligated DNA extract from David and Peng.
 
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*Mixed 6 µL insert (E0241), 2 µL vector (R0010), 2 µL DNA dilution buffer in reaction vial
 
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*Added 10 µL ligation buffer
 
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*Added 1 µL ligase, mixed
 
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*Let incubate 5min at room temperature
 
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'''Transformation'''
 
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*Mixed 20 µL of ligation product with 30 µL competent cells
 
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*Negative control: 2 µL of dH20 with 30 µL competent cells
 
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*Positive control: 5 µL of positive control with 30 µL competent cells
 
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*Incubate on ice for 20min
 
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*Heat shock @42C for 30s
 
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*Rest on ice for 2min
 
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*Incubate overnight @37C
 
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== 2006/06/14 ==
 
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'''Biobrick tutorial'''
 
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''More detailed procedure notes are in lab notebook.''
 
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*Began with 12 samples (3x each of negative control, R0010, E0241, E7104). Fortunately our negative controls showed no growth overnight.
 
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*Made 9 minipreps for the remaining samples. Set aside the E7104 samples (they are a positive control).
 
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*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.
 
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*Added Calf Intestinal Phosphatase (CIP) to R0010 samples to prevent self-ligation of the R0010 vector.
 
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*Performed gel electrophoresis on R0010 and E0241 samples.
 
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**Our e-gel was not entirely successful.
 
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[[Image:HH-JL biobrick-tutorial egel.jpg]]
 
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Must be more careful next time.
 
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'''Notes on using the nanodrop'''
 
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*Nanodrop measures concentration of DNA in small samples (1 µL). It is apparently not very accurate.
 
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*Clean nanodrop thoroughly with kimwipes and ethanol, before and after each use
 
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*On desktop, "ND-1000"->"Nucleic Acid"
 
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*Before each set of samples:
 
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**Calibrate nanodrop with drop of distilled water
 
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**Calibrate nanodrop with drop of background (solute), e.g. distilled water or elution buffer
 
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*Add sample to nanodrop, enter name of sample, "Measure"
 
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**Note: mix sample before placing on nanodrop
 

Revision as of 22:32, 16 June 2006

About

Undergraduate concentrating in Computer Science (class of 2007), member of 2006 Harvard iGem team.

Lab Notebook

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