Versions Compared

Old Version 85

changes.mady.by.user Unknown User (gary_9@mit.edu)

Saved on

compared with

New Version 86

changes.mady.by.user Unknown User (gary_9@mit.edu)

Saved on

Key

  • This line was added.
  • This line was removed.
  • Formatting was changed.

l

Toggle Cloak
SAMPLE

Cloak

Sample Experiment #1 (Template)

4 incomplete Day 1: Seed cells 5 incomplete Day 2: Transfect Promoter:Gene1 (___ ng/uL - transfection marker), Promoter:Gene2 (___ ng/uL) and Promoter:Gene3 (___ ng/uL) into HEK cells 8 incomplete Day 3: Add DOX (if necessary) 6 incomplete Day 4 (time): Flow Cytometer
  • DNA Dilution: __uL Promoter:Gene1; __uL Promoter:Gene2; __uL Promoter:Gene3
  • (Eg)x: add _:_:_
    1. __uL Promoter:Gene1-lipid complex
    2. __uL Promoter:Gene2-lipid complex
    3. __uL Promoter:Gene3-lipid complex


24-WELL PLATE TRANSFECTION MAP:

 

+ve control
  • Promoter:Gene1
+ve control
  • Promoter:Gene1
-ve control
  • dummy DNA
  • with Lipofectamine
-ve control
  • dummy DNA
  • with Lipofectamine
-ve/-ve control
  • no DNA
  • no Lipofectamine
-ve/-ve control
  • no DNA
  • no Lipofectamine
///EMPTY WELL/////////////////////////////////
////////////////////////////////////
(Eg)1
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)2
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)3
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)4
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)5
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3
(Eg)6
  • __uM DOX
  • Promoter:Gene1
  • Promoter:Gene2
  • Promoter:Gene3

 

RESULT: Brief description of experimental outcome (mean transfection efficiency and implications). Link to results page with data analysis.

Sample Experiment #2

(This was Practice Experiment #2)

36 complete 7/3: Seed cells 23 complete 7/4: Transfect hEF1a:eYFP (472 ng/uL) and hEF1a:tagBFP (1176 ng/uL) into HEK cells 24 complete 7/6 (1pm): Flow Cytometer

 

  • DNA Dilution: 10uL hEF1a:eYFP; 5uL hEF1a:tagBFP
  • (Y+B)x: add 1:1 (25uL hEF1a:eYFP/tagBFP-lipid complex)


Y1
  • hEF1a:eYFP
(Y+B)1
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve control
  • dummy DNA
  • with Lipofectamine
Y2
  • hEF1a:eYFP
(Y+B)2
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve control
  • dummy DNA
  • with Lipofectamine
B1
  • hEF1a:tagBFP
(Y+B)3
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve/-ve control
  • no DNA
  • no Lipofectamine
B2
  • hEF1a:tagBFP
(Y+B)4
  • hEF1a:eYFP
  • hEF1a:tagBFP
//////////////////-ve/-ve control
  • no DNA
  • no Lipofectamine


RESULT: Extremely low transfection efficiency (<10%). Repeat to improve co-transfection results.

...