User:Elaine Marie Robbins/Notebook/CHEM-496/2011/09/20: Difference between revisions
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==Objective== | ==Objective== | ||
To perform a PCR reaction to mutate green fluorescent | To perform a PCR reaction to mutate green fluorescent protein (GFP) to contain a cysteine amino acid just after the enterokinase cleavage site. | ||
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*Forward Primer: | *Forward Primer: | ||
::GAT AAG GAT GAC GAT AAG TGT CGA TGG GGA TCC GAA TTC GCC | ::5'- GAT AAG GAT GAC GAT AAG TGT CGA TGG GGA TCC GAA TTC GCC -3' | ||
*Reverse Primer: | *Reverse Primer: | ||
::GGC GAA TTC GGA TCC CCA TCG ACA CTT ATC GTC ATC CTT ATC | ::5'- GGC GAA TTC GGA TCC CCA TCG ACA CTT ATC GTC ATC CTT ATC -3' | ||
Line 48: | Line 48: | ||
*Forward Primer: | *Forward Primer: | ||
::GAC GAT GAC GAT AAG GAT CGA TGG GGA TCC GAA | ::5'- GAC GAT GAC GAT AAG GAT CGA TGG GGA TCC GAA -3' | ||
*Reverse Primer: | *Reverse Primer: | ||
::TTC GGA TCC CCA TCG ATC CTT ATC GTC ATC GTC | ::5'- TTC GGA TCC CCA TCG ATC CTT ATC GTC ATC GTC -3' | ||
==Notes== | ==Notes== |
Revision as of 10:25, 1 November 2011
Mutation and Amplification of GFP | <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> |
ObjectiveTo perform a PCR reaction to mutate green fluorescent protein (GFP) to contain a cysteine amino acid just after the enterokinase cleavage site.
Description
DataPrimer determined using http://www.basic.northwestern.edu/biotools/oligocalc.html:
Primer actually used in experiment:
NotesPrimers must have a melting point Tm ≥ 78°C, a GC content ≥ 40%, and be between 25-45 bases in length. Sequence of GFP vector provided by Invitrogen.
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