User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/04/04: Difference between revisions
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# Stop the gel when the yellow dye (Orange G) reaches the desired place on the gel (~1 hr.). | # Stop the gel when the yellow dye (Orange G) reaches the desired place on the gel (~1 hr.). | ||
# Remove the gel from the chamber and photograph under UV light. | # Remove the gel from the chamber and photograph under UV light. | ||
# Use a scalpel to cut the appropriate sized band(s) from the gel, place each gel slice in a 1.5 mL tube, and purify the DNA ( | # Use a scalpel to cut the appropriate sized band(s) from the gel, place each gel slice in a 1.5 mL tube, and purify the DNA with Zymoclean Gel DNA Recovery Kit(Follow the protocol inside the kit box). | ||
# Measure the concentration of the purified fragment samples with a Nanodrop Spectrophotometer. Record the absorbance (A260), purity (A260/A280), and concentration (ng/μl) for each sample. | # Measure the concentration of the purified fragment samples with a Nanodrop Spectrophotometer. Record the absorbance (A260), purity (A260/A280), and concentration (ng/μl) for each sample. | ||
[[Image:PcTF-RBS-Digestion-Gel-Behzad4-4-12.jpg|thumb|350px|White dashed lines border where the gel was cut to excise vector fragment (RBS) and insert fragment (PcTF).]] | |||
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[[category:OWWLabNotebookV1]] | [[category:OWWLabNotebookV1]] |
Revision as of 17:44, 4 April 2012
PcTF Subcloning in E. coli | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||
04/04/12General plan for assemblies
Assemblies
> Digests (Fermentas FD)
Separate the fragments via gel electrophoresis and purify the fragments
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