User:Matt Hartings/Notebook/Photosynthesis/2013/02/12

From OpenWetWare
Jump to navigationJump to search
Protein Re-engineering <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

DNA Gel

Maddie 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

  1. Ladder
  2. WT
  3. F1
  4. F2
  5. F3
  6. F4

(each lane consists of 5uL DNA and 1uL loading buffer).

Hb extraction

Last week we expressed WT Asc Mn-Hb and Ni-Hb. I am going to extract the Mn-Hb today.

  1. Sonicate: 30 seconds on, 30 seconds off, 3x
  2. balance in oak ridge tubes
  3. centrifuge for 2 hours