Haynes Lab:Notebook/Engineering PC-TFs/2014/10/13: Difference between revisions

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(Autocreate 2014/10/13 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
 
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==Summary==
==Summary==
*  
*  
'''Plate reader measurements'''
RFP signal
* Followed the fluorescent protein assay: http://openwetware.org/wiki/Haynes:ExtFluorProtein
* Used 100 μL per well
* Results:
# BL09 1 = 181
# BL09 2 (no transfection) = 25
# BL09 negative ctrl (no DNA) = 34
Bradford assay
* Followed Bradford Assay protocol: http://openwetware.org/wiki/Haynes:Bradford
* Used 5 μL for each BL09 sample
* OD 590 readings:
# Standard 1 (0 μg) = 0.276
# Standard 2 (1 μg) = 0.234
# Standard 3 (2 μg) = 0.394
# Standard 4 (4 μg) = 0.443
# Standard 5 (8 μg) = 0.644
# Standard 6 (16 μg) = 0.829
# BL09 1 = 0.31
# BL09 2 (no transfection) = 0.3
# BL09 negative = 0.047
Standard curve
[[Image:CG091514_Bradford.jpg|400px]]
Final results:
{| {{table}}
| Sample||Protein μg/μL||Protein*100 μL||Raw Fluor. (100 uL)||Fluor/ μg protein
|-
| BL09 1||10.42||1041.96||109||0.104610111
|-
| BL09 2||10.20||1019.64||610||0.598248687
|-
| BL09 negative||9.75||975.00||47||0.048205128
|}




Revision as of 15:41, 13 October 2014



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Summary

Plate reader measurements

RFP signal

  1. BL09 1 = 181
  2. BL09 2 (no transfection) = 25
  3. BL09 negative ctrl (no DNA) = 34


Bradford assay

  1. Standard 1 (0 μg) = 0.276
  2. Standard 2 (1 μg) = 0.234
  3. Standard 3 (2 μg) = 0.394
  4. Standard 4 (4 μg) = 0.443
  5. Standard 5 (8 μg) = 0.644
  6. Standard 6 (16 μg) = 0.829
  7. BL09 1 = 0.31
  8. BL09 2 (no transfection) = 0.3
  9. BL09 negative = 0.047

Standard curve


Final results:

Sample Protein μg/μL Protein*100 μL Raw Fluor. (100 uL) Fluor/ μg protein
BL09 1 10.42 1041.96 109 0.104610111
BL09 2 10.20 1019.64 610 0.598248687
BL09 negative 9.75 975.00 47 0.048205128



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