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

GMU:DIY Biolab “Driver’s License”/Marie Laure Ebel (view source)

Revision as of 21:49, 10 February 2018

253 bytes removed , 10 February 2018

→‎PRACTICING PCR ANALYSIS

Miga

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 ===Step 1. Extracting DNA from blood and from pork liver meat===
+<gallery>
-[[File:STEP1.1.jpg|170px]]
+File:STEP1.1.jpg
-[[File:STEP1.2.jpg|170px]]
+File:STEP1.2.jpg
-[[File:STEP1.3.jpg|170px]]
+File:STEP1.3.jpg
-[[File:STEP1.4.jpg|170px]]
+File:STEP1.4.jpg
-[[File:STEP1.5.jpg|170px]]
+File:STEP1.5.jpg
-[[File:STEP1.6.jpg|170px]]
+File:STEP1.6.jpg
-[[File:STEP1.7.jpg|170px]]
+File:STEP1.7.jpg
-[[File:STEP1.9.jpg|170px]]
+File:STEP1.9.jpg
-[[File:STEP1.10.jpg|170px]]
+File:STEP1.10.jpg
-[[File:STEP1.11.jpg|170px]]
+File:STEP1.11.jpg
-[[File:STEP1.8.jpg|170px]]
+File:STEP1.8.jpg
+</gallery>
 ===Step 2. STR-analysis with pork meat primer===
+<gallery>
-[[File:STEP 2.1.jpg|170px]]
+File:STEP 2.1.jpg
-[[File:STEP 2.2.jpg|170px]]
+File:STEP 2.2.jpg
-[[File:STEP2.3.jpg|170px]]
+File:STEP2.3.jpg
-[[File:STEP 2.4.jpg|170px]]
+File:STEP 2.4.jpg
-[[File:STEP2.5.jpg|170px]]
+File:STEP2.5.jpg
-[[File:STEP 2.6.jpg|170px]]
+File:STEP 2.6.jpg
-[[File:STEP 2.7.jpg|170px]]
+File:STEP 2.7.jpg
+</gallery>
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 ====#First attempt====
+<gallery>
+File:STEP 3.1.JPG
+File:STEP3.2.JPG
+File:STEP3.3.JPG
+File:STEP 3.4.jpg
+File:STEP 3.5.jpg
+File:STEP 3.6.jpg
+File:STEP 3.7.JPG
+File:STEP3.8.JPG
+File:STEP3.9.JPG
+File:STEP3.10.JPG
+</gallery>
-[[File:STEP 3.1.JPG|170px]]
+I got no results: either the ladder nor the DNA samples were "growing" as they should (multiple lines).
-[[File:STEP3.2.JPG|170px]]
-[[File:STEP3.3.JPG|170px]]
-[[File:STEP 3.4.jpg|170px]]
-[[File:STEP 3.5.jpg|170px]]
-[[File:STEP 3.6.jpg|170px]]
-[[File:STEP 3.7.JPG|170px]]
-[[File:STEP3.8.JPG|170px]]
-[[File:STEP3.9.JPG|170px]]
-[[File:STEP3.10.JPG|170px]]
-=> 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 with the samples themselves or with the preparation (agarose + buffer TAE 50x+ distilled water) => #Second attempt
 ====#Second attempt====
+<gallery>
+File:STEP 3.2.1.jpg
+File:STEP 3.2.2.jpg
+File:STEP 3.2.3.jpg
+File:STEP 3.2.4.jpg
+</gallery>
-[[File:STEP 3.2.1.jpg|200px]]
+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 quite old). Then, I noticed that the positive side was turning in blue color and some part of the agarose gel was melting down.
-[[File:STEP 3.2.2.jpg|200px]]
-[[File:STEP 3.2.3.jpg|200px]]
-[[File:STEP 3.2.4.jpg|200px]]
-=> 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 quite 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 blue 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====
+<gallery>
+File:STEP 3.3.1.jpg
+File:STEP 3.3.2.jpg
+</gallery>
-[[File:STEP 3.3.1.jpg|400px]]
+Still turning blue. Even with smaller electrodes that are completely covered by the solution (buffer TAE 50x + distilled water)
-[[File:STEP 3.3.2.jpg|400px]]
-=> Still turning blue. 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====
+<gallery>
+File:STEP 3.4.1.jpg
+File:STEP 3.4.2.jpg
+</gallery>
-[[File:STEP 3.4.1.jpg|400px]]
+First picture: distilled water only. Second picture: distilled water + buffer TAE 50x. No changes at all.
-[[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 the electrophoresis itself.
+To resolve this problem, I have asked different professional/scientist for help in that matter. In addition, I am also searching for protocols which allow determining human DNA above animals/plants.
->> To resolve this problem, I have asked different professional/scientist for help in that matter. In addition, I am also searching for protocols which allow determining human DNA above animals/plants.
-References:
+====References====
 [https://isogg.org/wiki/Admixture_analyses#AncestryDNA_-_Genetic_Ethnicity]
 [https://isogg.org/wiki/Autosomal_DNA#Autosomal_DNA_for_beginners]