GMU:DIY Biolab “Driver’s License”/Marie Laure Ebel: Difference between revisions

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[[File:STEP 3.2.4.jpg|400px]]
[[File:STEP 3.2.4.jpg|400px]]


=> Same problem : the ladder was growing as one bloc only (see the little drawing) + we noticed that the positive side was turning in blue color and some part of the agarose gel was melting down.
=> Same problem : the ladder was growing as one bloc only (see the little drawing). Even during a 2.2 attempt with new SERVA Stain G (which I thought it could be the problem, since the previous tube was quiet old). Then, I noticed that the positive side was turning in blue color and some part of the agarose gel was melting down.  
So we decided to test if this bleu color is coming from oxidation of the electrodes themselves, or if the ladder was "swimming" on the extra amount of solution (buffer TAE 50x + distilled water) above the agarose to reach the positive side => #Third attempt
So I decided to test if this bleu color is coming from oxidation of the electrodes themselves, or if the ladder was "swimming" on the extra amount of solution (buffer TAE 50x + distilled water) above the agarose to reach the positive side => #Third attempt




#Third attempt
#Third attempt
[[File:STEP 3.3.1.jpg|400px]]
[[File:STEP 3.3.2.jpg|400px]]
=> Still turning bleu. Even with smaller electrodes that are completely covered by the solution (buffer TAE 50x + distilled water)
So I decided to test if this oxidation is coming from the buffer or if it is simply a natural process => #Fourth attempt
#Fourth attempt
[[File:STEP 3.4.1.jpg|400px]]
[[File:STEP 3.4.2.jpg|400px]]
=> First picture : distilled water only. Second picture : distilled water + buffer TAE 50x. No changes at all.
So it might come from the SERVA Stain G (which is yellow-ish) or it is actually not a problem and does not affect on the electrophoresis itself.

Revision as of 19:26, 17 January 2018

// HUMAN DNA ANALYSIS //

Blood collected2.jpg DNA online analysis.png DND test technics.png DND test technics2.png DNA test technics3.png DND test limits.png DND test limits2.png DND testing sofware.png 11.08 DND analysis1.JPG 11.08 DND analysis2.JPG 11.08 DND analysis3.JPG [1] [2]


// PRACTICING PCR ANALYSIS //

Step 1. Extracting DNA from blood and from pork liver meat

STEP 1.1.jpg STEP 1.2.jpg STEP 1.3.jpg STEP 1.4.jpg STEP 1.5.jpg STEP 1.6.jpg STEP 1.7.jpg STEP 1.8.jpg STEP 1.9.jpg STEP 1.10.jpg STEP 1.11.jpg


Step 2. STR-analysis with pork meat primer

STEP 2.1.jpg STEP 2.2.jpg STEP 2.3.jpg STEP 2.4.jpg STEP 2.5.jpg STEP 2.6.jpg STEP 2.7.jpg


Step 3. Electrophoresis

  1. First attempt

STEP 3.1.JPG STEP 3.2.JPG STEP 3.3.JPG STEP 3.4.jpg STEP 3.5.jpg STEP 3.6.jpg STEP 3.7.JPG STEP 3.8.JPG STEP 3.9.JPG STEP 3.10.JPG

=> I got no results : either the ladder nor the DNA samples were "growing" as they should (multiple lines). So I decided to test the ladder only to see if there is a problem from the samples themselves or with the preparation (agarose + buffer TAE 50x+ distilled water) => #Second attempt

  1. Second attempt

STEP 3.2.1.jpg STEP 3.2.2.jpg STEP 3.2.3.jpg STEP 3.2.4.jpg

=> Same problem : the ladder was growing as one bloc only (see the little drawing). Even during a 2.2 attempt with new SERVA Stain G (which I thought it could be the problem, since the previous tube was quiet old). Then, I noticed that the positive side was turning in blue color and some part of the agarose gel was melting down. So I decided to test if this bleu color is coming from oxidation of the electrodes themselves, or if the ladder was "swimming" on the extra amount of solution (buffer TAE 50x + distilled water) above the agarose to reach the positive side => #Third attempt


  1. Third attempt

STEP 3.3.1.jpg STEP 3.3.2.jpg

=> Still turning bleu. Even with smaller electrodes that are completely covered by the solution (buffer TAE 50x + distilled water) So I decided to test if this oxidation is coming from the buffer or if it is simply a natural process => #Fourth attempt

  1. Fourth attempt

STEP 3.4.1.jpg STEP 3.4.2.jpg

=> First picture : distilled water only. Second picture : distilled water + buffer TAE 50x. No changes at all. So it might come from the SERVA Stain G (which is yellow-ish) or it is actually not a problem and does not affect on the electrophoresis itself.