User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/04/06: Difference between revisions
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# 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). | # 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 Biotek Take3 plate from the Stabenfeldt lab. Record the absorbance (A260), purity (A260/A280), and concentration (ng/μl) for each sample. | # 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. | ||
* Insert RBS-PcTF + (S/X) : 15 + 1123 + 6 = 1144 | |||
* Vector T7 strong promoter (BBa_I712074) 46 bp plasmid backbone is pSB1AK8 size: 3426 bp → 3426 + 46 = 3472 | |||
* Vector T7 promoter (BBa_I719005) 23 bp plasmid backbone is pSB1A2 size: 2079 bp → 2079 + 23 = 2102 | |||
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Revision as of 14:12, 7 April 2012
PcTF Subcloning in E. coli | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||
06/04/12Assemblies
> Digests (Fermentas FD)
Separate the fragments via gel electrophoresis and purify the fragments
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