User:Eric Ma/Notebook/MICB323 Lab Book/2009/02/10: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 39: Line 39:
# Mass Ruler (10 µL)
# Mass Ruler (10 µL)
Gel was run for 1 hour @ 120 volts. <b>Noticed that the samples in lanes 3, 5 and 7 had a tendency to float off.</b>
Gel was run for 1 hour @ 120 volts. <b>Noticed that the samples in lanes 3, 5 and 7 had a tendency to float off.</b>
==Part 3: Determination of Viral Lysate Protein Concentrations using the Bradford and Bicinchoninic Acid Protein Assays==




Revision as of 15:21, 10 February 2009

Project name <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>

Part 1: Agarose Gel of Plasmid DNA from Previous Lab

  • Conditions Used:
    • 100 mL of 0.7% agarose gel + 10µL SYBR Safe Stain

Summary of Tube Labels

    • EM1-4 EcoRI - Manipulation of EcoRI concentration for digestion of isolated pDNA
    • EM1-4 NaCl - Manipulation of NaCl concentration for digestion of isolated pDNA
    • EM#1-2 - Isolated plasmid DNA from CaCl2 (#1: transformed) and (#2: untransformed)
    • EM1-3 - Digested plasmid DNA from CaCl2 (#1: transformed), (#2: untransformed),(#3: Sunny's Part 2)


Lanes on the gel and volumes added to the wells

  1. Supercoiled Ladder (10µL)
  2. 380 and 1000 standard (10µL)
  3. EM1 - Transformed and digested pDNA from CaCl2 (10 µL Sample + 2µL 6X Loading Buffer)
  4. EM#1 - Isolated transformed pDNA from CaCl2 (10 µL Sample + 2µL 6X Loading Buffer)
  5. EM2 - Untransformed and digested pDNA from CaCl2 (10 µL Sample + 2µL 6X Loading Buffer)
  6. EM#2 - Isolated untransformed pDNA from CaCl2 (10 µL Sample + 2µL 6X Loading Buffer)
  7. EM3 - Isolated p421 plasmid DNA inserted via CaCl2 (10 µL Sample + 2µL 6X Loading Buffer) (Note: From Sunny)
  8. EM#3 - Digested p421 plasmid DNA inserted via CaCl2 (10 µL Sample + 2µL 6X Loading Buffer) (Note: From Sunny)
  9. Ligation Reaction (8µL including 1.5µL of 6X Loading Buffer)
  10. Mass Ruler (10 µL)
  11. EM1 EcoRI (18 µL)
  12. EM2 EcoRI (18 µL)
  13. EM3 EcoRI (18 µL)
  14. EM4 EcoRI (18 µL)
  15. EM1 NaCl (18 µL)
  16. EM2 NaCl (18 µL)
  17. EM3 NaCl (18 µL)
  18. EM4 NaCl (18 µL)
  19. Mass Ruler (10 µL)

Gel was run for 1 hour @ 120 volts. Noticed that the samples in lanes 3, 5 and 7 had a tendency to float off.

Part 3: Determination of Viral Lysate Protein Concentrations using the Bradford and Bicinchoninic Acid Protein Assays


Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:Eric_Ma/Notebook/MICB323_Lab_Book/2009/02/10&oldid=284729"