User:Matt Hartings/Notebook/Photosynthesis/2013/02/12: Difference between revisions
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Last week we expressed WT Asc Mn-Hb and Ni-Hb. I am going to extract the Mn-Hb today. | Last week we expressed WT Asc Mn-Hb and Ni-Hb. I am going to extract the Mn-Hb today. | ||
# Sonicate: 30 seconds on, 30 seconds off, 3x | # Sonicate: 30 seconds on, 30 seconds off, 3x | ||
# balance in oak ridge tubes | # balance in oak ridge tubes to within 0.01g | ||
# centrifuge for 2 hours | # centrifuge for 2 hours, 4C, at 18000rpm | ||
Nyousha is making buffers for FPLC: 1L of 10mM Tris 50mM NaCl pH 8.5, 1L of 10mM Tris 1M NaCl pH 8.5 | |||
The buffers were filtered after they were made. | |||
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Revision as of 13:07, 12 February 2013
 Protein Re-engineering
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<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> |
DNA GelMaddie ran several PCR runs last week to try to mutate K31C Asc Hb to K31C T109C Asc Hb. I made an agarose gel (1% - 0.25g agarose in 25mL TAE Buffer) The lanes in the gel are
(each lane consists of 5uL DNA and 1uL loading buffer). Results Lane 2 showed nothing. Lanes 3-6 were really streaky. Potentially many different products, or perhaps too concentrated. Should run again more dilute. Hb extractionLast week we expressed WT Asc Mn-Hb and Ni-Hb. I am going to extract the Mn-Hb today.
Nyousha is making buffers for FPLC: 1L of 10mM Tris 50mM NaCl pH 8.5, 1L of 10mM Tris 1M NaCl pH 8.5 The buffers were filtered after they were made. | |
