User:Elizabeth Ghias/Notebook/Experimental Chemistry/2013/03/27

From OpenWetWare

< User:Elizabeth Ghias | Notebook | Experimental Chemistry | 2013 | 03(Difference between revisions)
Jump to: navigation, search
Current revision (11:26, 3 April 2013) (view source)
 
(2 intermediate revisions not shown.)
Line 15: Line 15:
==Description==
==Description==
-
# The hydrogels were removed from the freezer at 10 am and were put back in at 2 pm.
+
* The hydrogels were removed from the freezer at 10 am and were put back in at 2 pm.
-
# The quaternary ammonium salt films made on monday could not be crosslinked because they had not set.
+
* The quaternary ammonium salt films made on monday could not be crosslinked because they had not set.
-
# The 10% QAS film was filtered using a vacuum pump and a buchner funnel.
+
* The 10% QAS film was filtered using a vacuum pump and a buchner funnel.
 +
 
<u>Quaternary Ammonium Salt Film Preparation</u>
<u>Quaternary Ammonium Salt Film Preparation</u>
 +
 +
# Measure out two 1 g samples of the 1% QAS
 +
# Place in beaker with 30 mL water.  Dissolve on heat.
 +
# Once dissolved, add 1 mL glutaraldehyde.  Allow to stir for 3 minutes.
 +
# Pour into plastic dish and allow to evaporate until Friday.
 +
 +
 +
<u>Next Hydrogel Reaction (Click Chemistry)</u>
 +
 +
* PNAS article <u>A chemical method for fast and sensitive detection of DNA synthesis in vivo</u> by Adrian Salic and Timothy J. Mitchison was used to determine the ranges of concentrations for the reactants for the click reaction.
 +
* The article uses 0.5 - 1 mM CuSO<sub>4</sub>, 1 - 100 μM fluorescent azide (in 10 - 100 mM DMSO stock), and 50 - 100 mM ascorbic acid (added last from a stock solution of 0.5 M)
 +
* They reacted for 30 minutes at room temperature
 +
* The dye should be in a solution of DMSO
 +
* We will add 1.5X excess of dye
 +
* Will use the same concentration of CuSO<sub>4</sub> and ascorbic acid as papers (so depends on however much water we decide to add)
==Data==
==Data==
-
* Add data and results here...
 
-
==Notes==
+
No data was collected today.
 +
==Notes==
-
<u>Schedule</u>
+
* Next step is to figure out the amount of alkyne (assuming no loss) and then calculate the rest of the concentrations
-
Friday - crosslink the rest of the QAS films
 
-
Monday - make bacteria starter culture
 
-
Tuesday - plate the bacteria on the films
 
-
Wednesday - read the films
 

Current revision

Experimental Chemistry Main project page
Previous entry      Next entry


Objective

Take out the hydrogels to thaw, crosslink the quaternary ammonium salt films made monday (0.01%), make 1% QAS film, filter the 10% QAS, and look up concentrations for the next hydrogel reaction.

Description

  • The hydrogels were removed from the freezer at 10 am and were put back in at 2 pm.
  • The quaternary ammonium salt films made on monday could not be crosslinked because they had not set.
  • The 10% QAS film was filtered using a vacuum pump and a buchner funnel.


Quaternary Ammonium Salt Film Preparation

  1. Measure out two 1 g samples of the 1% QAS
  2. Place in beaker with 30 mL water. Dissolve on heat.
  3. Once dissolved, add 1 mL glutaraldehyde. Allow to stir for 3 minutes.
  4. Pour into plastic dish and allow to evaporate until Friday.


Next Hydrogel Reaction (Click Chemistry)

  • PNAS article A chemical method for fast and sensitive detection of DNA synthesis in vivo by Adrian Salic and Timothy J. Mitchison was used to determine the ranges of concentrations for the reactants for the click reaction.
  • The article uses 0.5 - 1 mM CuSO4, 1 - 100 μM fluorescent azide (in 10 - 100 mM DMSO stock), and 50 - 100 mM ascorbic acid (added last from a stock solution of 0.5 M)
  • They reacted for 30 minutes at room temperature
  • The dye should be in a solution of DMSO
  • We will add 1.5X excess of dye
  • Will use the same concentration of CuSO4 and ascorbic acid as papers (so depends on however much water we decide to add)


Data

No data was collected today.

Notes

  • Next step is to figure out the amount of alkyne (assuming no loss) and then calculate the rest of the concentrations




Personal tools