Difference between revisions of "GDSYZX-United/PROJECT/extract dna"

 
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<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/extract _dna">Extract DNA in arabidopsis thaliana</a></li>
 
<li><a href="https://2016.igem.org/GDSYZX-United/PROJECT/enzyme_cleavage">Enzyme cleavage</a></li>
 
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<p><div class="text_font"><strong>Place:</strong>Biology Experiment Teaching Center Of Sun Yat-sen University</div></p>
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<p><div class="text_font" style="text-indent:0em;"><strong>Place:</strong>Biology Experiment Teaching Center Of Sun Yat-sen University</div></p>
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<p><div class="text_font"><strong>Date:</strong>July 25th, 2016</div></p>
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<p><div class="text_font"><strong>Experimental apparatus:</strong>Grinding rods, 2 ml centrifuge tube, double panel, transfer liquid gun, adsorption column &collection tube and centrifuge.</div></p>
 
<p><div class="text_font"><strong>Experimental apparatus:</strong>Grinding rods, 2 ml centrifuge tube, double panel, transfer liquid gun, adsorption column &collection tube and centrifuge.</div></p>
 
<p><div class="text_font"><strong>Experimental drugs:</strong>Buffer PCB, β- mercaptoethanol, RNaseA, chloroform, Buffer BD, anhydrous ethanol, PW Solution, Wash Solution, TE Buffer.</div></p>
 
<p><div class="text_font"><strong>Experimental drugs:</strong>Buffer PCB, β- mercaptoethanol, RNaseA, chloroform, Buffer BD, anhydrous ethanol, PW Solution, Wash Solution, TE Buffer.</div></p>
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<li>Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.</li>
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<li style="text-indent:0em;">Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.</li>
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<li>There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.</li>
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<li style="text-indent:0em;">There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.</li>
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<li>Not all reagents were fully blended.</li>
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<li style="text-indent:0em;">Not all reagents were fully blended.</li>
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<li>The usage of transfer liquid gun was not normative.</li>
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<li style="text-indent:0em;">The usage of transfer liquid gun was not normative.</li>
 
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<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/f/f5/T--GDSYZX-United--project_extract_dna5.jpg">
 
<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/f/f5/T--GDSYZX-United--project_extract_dna5.jpg">
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<p><div class="text_font">result</div></p>
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<p><div class="text_font">result(we falied this time)</div></p>
 
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Latest revision as of 14:51, 17 October 2016

Extract DNA in arabidopsis thaliana

Place:Biology Experiment Teaching Center Of Sun Yat-sen University

Experimental apparatus:Grinding rods, 2 ml centrifuge tube, double panel, transfer liquid gun, adsorption column &collection tube and centrifuge.

Experimental drugs:Buffer PCB, β- mercaptoethanol, RNaseA, chloroform, Buffer BD, anhydrous ethanol, PW Solution, Wash Solution, TE Buffer.

Procedure:

Experimental drugs operation Time
50-100mg Young leaves of Arabidopsis thaliana. Transfer to 2 ml centrifuge tube after grinding with grinding bar.
600μl 65℃ Buffer PCB & 12μl β- mercaptoethanol Placed in 65 ℃ water after blending. 25min
20μl RNaseA waiting 5min
About 630μl chloroform Transfer supernatant liquid to a clean 2 ml centrifuge tube after blending and following 12000 RPM centrifuge. centrifuge：5min
About 400μl Buffer BD Blending for 3-5 times.
About 400μl anhydrous ethanol Transfer to the adsorption column after blending,and then waiting. Dump the waste liquid after centrifuging with 10000 RPM. waiting：2min
centrifuging：1min
500μl Wash Solution Centrifuging with 10000 RPM after adding the reagent, and then dump the waste liquid. 1min
centrifuge with 12000 RPM. 2min
50μl TE Buffer Place the adsorption column in the new 2 ml centrifuge tube, then waiting after adding reagent. wait:3min
centrifuge with 12000 RPM. 2min

Result:

The target DNA can be extracted successfully in most of the groups, but they have very low concentration and many impurities.

Introspection:

  1. Understanding of the experimental requirements was not very well and had some wrong operations in the experiment.
  2. There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.
  3. Not all reagents were fully blended.
  4. The usage of transfer liquid gun was not normative.

image

arabidopsis thaliana

image

grinding with grinding bar

image

After blending with Buffer PCB

image

centrifuge

image

result(we falied this time)