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									<h1><strong>Extract DNA in arabidopsis thaliana</strong></h1>	
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									<p><div class="text_font"><strong>Place:</strong>Biology Experiment Teaching Center Of Sun Yat-sen University</div></p>
									<p><div class="text_font"><strong>Date:</strong>July 25th, 2016</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>
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									<p><div class="text_font"><strong>Procedure:</strong></div></p>	
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												<th>Experimental drugs</th>
												<th>operation</th>
												<th>Time</th>
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												<td>50-100mg Young leaves of Arabidopsis thaliana.</td>
												<td>Transfer to 2 ml centrifuge tube after grinding with grinding bar.</td>
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												<td>600μl 65℃ Buffer PCB  & 12μl β- mercaptoethanol</td>
												<td>Placed in 65 ℃ water after blending.</td>
												<td>25min</td>
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												<td>20μl RNaseA</td>
												<td>waiting</td>
												<td>5min</td>
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												<td>About 630μl chloroform</td>
												<td>Transfer  supernatant liquid to a clean 2 ml centrifuge tube after blending and following 12000 RPM centrifuge.</td>
												<td>centrifuge：5min</td>
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												<td>About 400μl Buffer BD</td>
												<td>Blending for 3-5 times. </td>
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												<td>About 400μl anhydrous ethanol</td>
												<td>Transfer to the adsorption column after blending,and then waiting. Dump the waste liquid after centrifuging with 10000 RPM.</td>
												<td>waiting：2min<br>centrifuging：1min</td>
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												<td>500μl Wash Solution</td>
												<td>Centrifuging with 10000 RPM after adding the reagent, and then dump  the waste liquid.</td>
												<td>1min</td>
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												<td>centrifuge with 12000 RPM.</td>
												<td>2min</td>
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												<td>50μl  TE Buffer</td>
												<td>Place the adsorption column in the new 2 ml centrifuge tube, then waiting after adding reagent.</td>
												<td>wait:3min</td>
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												<td>centrifuge with 12000 RPM.</td>
												<td>2min</td>
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									<p><div class="text_font"><strong>Result:</strong></div></p>
									<p><div class="text_font">The target DNA can be extracted successfully in most of the groups, but they have very low concentration and many impurities.</div></p>
									<p><div class="text_font"><strong>Introspection:</strong></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>
											<li>There is still many liquid residue on the grinding rod after grinding Arabidopsis thaliana leaves.</li>
											<li>Not all reagents were fully blended.</li>
											<li>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/2016/9/94/T--GDSYZX-United--project_extract_dna1.jpg">
										<p><div class="text_font">arabidopsis thaliana</div></p>
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											<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/8/80/T--GDSYZX-United--project_extract_dna2.jpg">
											<p><div class="text_font">grinding with grinding bar</div></p>
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											<img alt="image" class="img-responsive" src="https://static.igem.org/mediawiki/2016/b/b1/T--GDSYZX-United--project_extract_dna3.jpg">
											<p><div class="text_font">After blending with  Buffer PCB</div></p>
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											<p><div class="text_font">centrifuge</div></p>
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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">
											<p><div class="text_font">result</div></p>
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