<p class="jove_title" style="text-align: justify">白蚁的整个肠道DNA提取的程序总结为： </p><ol><li>冰浴白蚁，清除肠道使用无菌镊子和稳定肠道样本缓冲液中。 </li><li> PVPP / SDS /酚​​缓冲区均质样品。 </li><li>使用Qiagen公司DNeasy列从原油裂解的DNA提取和纯化。 </li></ol><p class="jove_title" style="text-align: justify">协议： </p><ol><li>在冰上，消除白蚁用无菌镊子从工人等级的胆量。 </li><li>立即转移的胆量和内容，无菌，无核酸管内含有50微升冰冷1X分子生物学级TE缓冲液（1毫米的Tris - HCl，0.1 mM的EDTA，pH值8.0）。冻结在-20 ° C的样本，或进行直接与同质化。 </li><li>肠道样品和缓冲传输无菌，无核酸酶2毫升螺杆上限管预装500毫克无菌氧化锆/二氧化硅珠（0.1毫米）和700μl1X TE缓冲液液，含有1％W / V polyvinylpolypyrrolidone（PVPP ）。 </li><li>加入50μL20％十二烷基硫酸钠（SDS），苯酚样品和500μL。 </li><li>最高的设置使用三个周期同质化的30秒和30秒冰寒蝉均质（珠节拍）。 </li><li>泥沙不溶性物质，在<em>8000</em> X克1分钟。 </li><li>净化的水层（最上层）原油裂解净化描述的方法使用Qiagen公司DNeasy列300μL等份。 </li><li>量化核酸含量在-20 ° C和冻结样品，以备后用。 </li></ol>

<ol>
	<li>Berthelet, M., Whyte, L. G., Greer, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid,+Direct+Extraction+of+DNA+from+Soils+for+PCR+Analysis+using+Polyvinylpolypyrrolidone+Spin+Columns.">Rapid, Direct Extraction of DNA from Soils for PCR Analysis using Polyvinylpolypyrrolidone Spin Columns.</a> <em style='font-style: italic'>FEMS Microbiol. Lett</em>. <b>138</b>, 17-22 (1996).</li><li>Kudo, T., Ohkuma, M., Moriya, S., Noda, S., Ohtoko, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+Phylogenetic+Identification+of+the+Intestinal+Anaerobic+Microbial+Community+in+the+Hindgut+of+the+Termite,+Reticulitermes+Speratus,+without+Cultivation.">Molecular Phylogenetic Identification of the Intestinal Anaerobic Microbial Community in the Hindgut of the Termite, Reticulitermes Speratus, without Cultivation.</a> <em style='font-style: italic'>Extremophiles</em>. <b>2</b>, 155-161 (1998).</li><li>Mor&#233;, M. I., Herrick, J. B., Silva, M. C., Ghiorse, W. C., Madsen, E. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+Cell+Lysis+of+Indigenous+Microorganisms+and+Rapid+Extraction+of+Microbial+DNA+from+Sediment.">Quantitative Cell Lysis of Indigenous Microorganisms and Rapid Extraction of Microbial DNA from Sediment.</a> <em style='font-style: italic'>Appl. Environ. Microbiol</em>. <b>60</b>, 1572-1580 (1994).</li><li>Odelson, D. A., Breznak, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volatile+Fatty+Acid+Production+by+the+Hindgut+Microbiota+of+Xylophagous+Termites.">Volatile Fatty Acid Production by the Hindgut Microbiota of Xylophagous Termites.</a> <em style='font-style: italic'>Appl. Environ. Microbiol</em>. <b>45</b>, 1602-1613 (1983).</li><li>Ottesen, E. A., Hong, J. W., Quake, S. R., Leadbetter, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidic+Digital+PCR+Enables+Multigene+Analysis+of+individual+Environmental+Bacteria.">Microfluidic Digital PCR Enables Multigene Analysis of individual Environmental Bacteria.</a> <em style='font-style: italic'>Science</em>. <b>314</b>, 1464-1467 (2006).</li><li>Purdy, K. J., Embley, T. M., Takii, S., Newdell, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+Extraction+of+DNA+and+rRNA+from+Sediments+by+a+Novel+Hydroxyapatite+Spin-Column+Method.">Rapid Extraction of DNA and rRNA from Sediments by a Novel Hydroxyapatite Spin-Column Method.</a> <em style='font-style: italic'>Appl. Environ. Microbiol</em>. <b>62</b>, 3905-3907 (1996).</li><li>Salmassi, T. M., Leadbetter, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+Genes+of+Tetrahydrofolate-Dependent+Metabolism+from+Cultivated+Spirochaetes+and+the+Gut+Community+of+the+Termite+Zootermopsis+Angusticollis.">Analysis of Genes of Tetrahydrofolate-Dependent Metabolism from Cultivated Spirochaetes and the Gut Community of the Termite Zootermopsis Angusticollis.</a> <em style='font-style: italic'>Microbiology</em>. <b>149</b>, 2529-2537 (2003).</li><li>Schmitt-Wagner, D., Friedrich, M. W., Wagner, B., Brune, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phylogenetic+Diversity,+Abundance,+and+Axial+Distribution+of+Bacteria+in+the+Intestinal+Tract+of+Two+Soil-Feeding+Termites+(Cubitermes+spp).">Phylogenetic Diversity, Abundance, and Axial Distribution of Bacteria in the Intestinal Tract of Two Soil-Feeding Termites (Cubitermes spp).</a> <em style='font-style: italic'>Appl. Environ. Microbiol</em>. <b>69</b>, 6007-6017 (2003).</li><li>Tsai, Y. L., Olson, B. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+Method+for+Separation+of+Bacterial+DNA+from+Humic+Substances+in+Sediments+for+Polymerase+Chain+Reaction.">Rapid Method for Separation of Bacterial DNA from Humic Substances in Sediments for Polymerase Chain Reaction.</a> <em style='font-style: italic'>Appl. Environ. Microbiol</em>. <b>58</b>, 2292-2295 (1992).</li></ol>

<p>根据我们的经验，从木材喂养白蚁喜欢Zootermopis nevadensis的微生物群落中提取的DNA PCR模板充分一轮的提取和纯化后的纯。然而，一些白蚁，如乱抛垃圾的喂养和土壤哺育物种可能有较高浓度的腐殖酸在其肠道内容，可能需要额外的净化肠道微生物DNA。总DNA的产量从5 Z。nevadensis工人的胆量是在10-30微克的范围。白蚁明显小于或大于这个物种，可能需要更多或更少的标本，获得了类似的DNA量。 </p><p>这种方法可以很容...

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		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>PVPP/SDS/phenol</td>
<td>Buffer</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>homogeneization buffer</td>
</tr>
<tr>
<td>DNeasy Tissue Kit</td>
<td>Kit</td>
<td>Qiagen</td>
<td>77607</td>
<td>Used according to the protocol for isolation of genomic DNA from crude lysates (Appendix H, product manual version: July 2003)</td>
</tr>
<tr>
<td>TE</td>
<td>Buffer</td>
<td>Sigma</td>
<td>T9285</td>
<td>1x buffer (1 mM Tris-HCl, 0.1 mM EDTA, pH 8.0) from 100x concentrate</td>
</tr>
<tr>
<td>zirconia/silica beads </td>
<td>Supplies</td>
<td>BioSpec Products</td>
<td>11079101z</td>
<td>0.1 mm</td>
</tr>
<tr>
<td>PVPP</td>
<td>Reagent</td>
<td>Sigma</td>
<td>P6755</td>
<td>1% w/v polyvinylpolypyrrolidone prepared from dry reagent as a 1x suspension in TE buffer </td>
</tr>
<tr>
<td>Zootermopsis nevadensis</td>
<td>Animal</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Termites</td>
</tr>
<tr>
<td>SDS</td>
<td>Reagent</td>
<td>Sigma</td>
<td>L4390</td>
<td>Sodium dodecyl sulfate 20% soln. in water from dry reagent</td>
</tr>
<tr>
<td>Phenol</td>
<td>Reagent</td>
<td>Sigma</td>
<td>77607</td>
<td>TE-saturated, ~73%</td>
</tr>
<tr>
<td>MiniBeadbeater-8</td>
<td>Tool</td>
<td>BioSpec Products</td>
<td>963</td>
<td>&nbsp;</td>
<tr>
<td>BSS</td>
<td>Buffered Salt Solution, pH 7.2</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Formulation per liter: 2.5 g  K2HPO4, 1.0 g  KH2PO4, 1.6 g KCl, 1.4 g NaCl, 0.075 g CaCl, 1 g MgCl, and  10 mL of a 1M soln. of NaHCO3</td>
</tr>
<tr>
<td>AxioPlan-2</td>
<td>Microscope</td>
<td>Carl Zeiss, Inc. USA</td>
<td>&nbsp;</td>
<td>Outfitted with 40x objective, 1.6x optivar and 10x ocular lenses. Samples were viewed using phase contrast illumination</td>
</tr>		</tbody>
		</table>
		</div>
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extracting dna from the gut microbes of the termite (zootermopsis angusticollis) and visualizing gut microbes

<p class="jove_content">白蚁是在已知有能力只消耗木材为生少数动物。白蚁肠道道包含一个高密度和物种丰富的微生物种群，在木质纤维素降解助攻主要成醋酸，助长白蚁代谢的关键营养素（Odelson Breznak，1983年）。在这些微生物种群是细菌，产甲烷古菌，在一些白蚁（“低”），真核细胞原生动物。因此，白蚁是优秀的科研课题研究微生物物种和许多他们执行他们的主人的利益的生化功能之间的相互作用。白蚁胆量，以及参与各种生化过程的关键基因的微生物种群的物种组成已探讨利用分子生物学技术（工藤等人，1998年，施密特，瓦格纳等，2003; Salmassi利百特，2003年）。这些技术依赖于从白蚁肠道内环境的高品质核酸的提取和纯化。此影片中所描述的提取工艺是从环境样品（铁道部等，1994核酸的提取和纯化的开发协议的修改编译;。Berthelet等，1996;普尔蒂等，1996;。Salmassi利德贝特2003年。Ottesen等; 2006年），它产生的DNA为模板，利用聚合酶链反应（PCR）从白蚁肠材料。</p>

Watch this Scientific Journal Video about Extracting DNA from the Gut Microbes of the Termite Zootermopsis Angusticollis and Visualizing Gut Microbes at JoVE.com

Extracting DNA from the Gut Microbes of the Termite Zootermopsis Angusticollis and Visualizing Gut Microbes

<p class="jove_content">该视频演示了居住在白蚁后肠微生物物种中提取DNA的技术。湿挂载幻灯片，这是有益的肠道微生物群落的可视化的准备工作也说明，并通过参观物种丰富的肠道环境。</p>

从白蚁肠道微生物（Zootermopsis Angusticollis）中提取的DNA和可视化肠道微生物

Termites are among the few animals known to have the capacity to subsist solely by consuming wood.  The termite gut tract contains a dense and ...

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Extracting DNA from the Gut Microbes of the Termite (Zootermopsis Angusticollis) and Visualizing Gut Microbes

15.4K 次观看.  California Institute of Technology - Caltech. 该视频演示了居住在白蚁后肠微生物物种中提取DNA的技术。湿挂载幻灯片，这是有益的肠道微生物群落的可视化的准备工作也说明，并通过参观物种丰富的肠道环境。