User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/04/05: Difference between revisions
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# Using a sterile pipette tip, touch the bacterial streak (or pick up a single colony) and put the tip into the LB medium (bacterial end down). | # Using a sterile pipette tip, touch the bacterial streak (or pick up a single colony) and put the tip into the LB medium (bacterial end down). | ||
# Grow the cultures for 7 hours in a shaking 37°C incubator.<br><br> | # Grow the cultures for 7 hours in a shaking 37°C incubator.<br><br> | ||
''Extract the plasmid DNA: Qiagen Miniprep Kit''' ''1.5 hours''<br> | |||
To extract the plasmid DNA from the bacteria, perform a mini prep (refer to the Qiagen miniprep protocol). 2 ml of culture usually gives a yield of about 200 ng/μl (elution vol. = 75 μl). Measure the concentration of the purified fragment samples with a Biotek Take3 plate from the Stabenfeldt lab. Record the absorbance (A260), purity (A260/A280), and concentration (ng/μl) for each sample.<br> | |||
Measure the concentration of the purified fragment samples with a Biotek Take3 plate from the Stabenfeldt lab. Record the absorbance (A260), purity (A260/A280), and concentration (ng/μl) for each sample. | |||
Biotek Take3 Result: | |||
# BBa_I712074 55.834 (ng/μl) , 2.083 (260/280) | |||
# BBa_I719005 (Re-transformed gene) 55.422 (ng/μl) , 2.115 (260/280) | |||
# BBa_B0030 (Re-transformed gene) 35.328 (ng/μl) , 2.114 (260/280) | |||
# BD111 (Constructed gene)-Colony A , 69.251 (ng/μl) , 2.091 (260/280) | |||
# BD111 (Constructed gene)-Colony B , 64.353 (ng/μl) , 2.085 (260/280) |
Revision as of 19:34, 5 April 2012
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5/4/2012Culturing the colonies grew on agar plates on 4/4/2012
Grow liquid cultures
Extract the plasmid DNA: Qiagen Miniprep Kit' 1.5 hours Measure the concentration of the purified fragment samples with a Biotek Take3 plate from the Stabenfeldt lab. Record the absorbance (A260), purity (A260/A280), and concentration (ng/μl) for each sample. Biotek Take3 Result:
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