Name: an introduction to worm lab: from culturing worms to mutagenesis
Uploaded: 2011-01-11T22:00:00
Description: Watch this Scientific Journal Video about An Introduction to Worm Lab: from Culturing Worms to Mutagenesis at JoVE.com

这项工作已经由美国国家科学基金会资助的IOB＃0615996。

第1部分：准备线虫的生长介质（NGM）的Petri板实验线虫， 如线虫和 P. pacificus通常在6厘米的Petri板含有线虫的生长介质（NGM）1实验室培养。蠕虫是保持在20 ° C，但可以根据线虫的物种和实验设计，在一定温度范围内（4 ° C - 25 ° C之间）孵育。为了准备与NGM板，标准的无菌技术已被用来防止真菌和细菌的污染。以下是协议准备NGM板： <ol><li>称取蛋...

<ol>
	<li>Brenner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Genetics. 77 (1), 71-71 (1974).</li><li>Anderson, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods Cell Biol. 48, 31-31 (1995).</li><li>Yandell, M. D., Edgar, L. G., Wood, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Proceedings of the National Academy of Sciences of the United States of America. 91 (4), 1381-1381 (1994).</li><li>Pires-daSilva, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> WormBook. , 1-1 (2005).</li><li>Kenning, C., Kipping, I., Sommer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> J. Genesis. 40 (3), 176-176 (2004).</li><li>Sommer, R. J., Carta, L. K., Kim, S. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fundamental and Applied Nematology. 19 (6), 511-511 (1996).</li><li>Pires-daSilva, A., Sommer, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Genes &#38; development. 18 (10), 1198-1198 (2004).</li><li>Wei, A., Yuan, A., Fawcett, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Nucleic acids research. 30 (20), 110-110 (2002).</li><li>Jorgensen, E. M., Mango, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Nat Rev Genet. 3 (5), 356-356 (2002).</li></ol>

突变是一个广泛采用的技术，在各种实验模式生物基因研究。通常用于突变实验，以确定一个基因的2的功能。这里所描述的突变程序可以使用，不仅为体育pacificus，但也为C。线虫和其他相关线虫物种。在这里，我们介绍两种方法，环境管理体系和TMP /紫外线诱变。 EMS是一种化学物质，诱导点突变或整个基因组的3,8小缺失。补骨脂突变（TMP / UV）...

an introduction to worm lab: from culturing worms to mutagenesis

这个协议描述的程序，以保持线虫在实验室，以及如何mutagenize他们使用两种可供选择的的方法：甲基磺酸乙酯（EMS）和4，5“，8 trimethylpsoralen结合紫外线（TMP /紫外线）。线虫是因为其简单的身体计划和交配系统，这是自我施肥雌雄同体和男性可以产生数以百计的每动物后代组成强大的生物遗传学的研究系统。线虫是保持中含有的细菌草坪的琼脂板，可以很容易地从一个板块转移到另一台使用一挑。 EMS是常用的一种烷化剂诱发点突变和小缺失，而TMP /紫外线主要是诱导删除。根据正在使用的线虫的物种，环境管理体系和TMP的含量进行优化。为了隔离隐性突变线虫Pristionchus pacificus，F2代的动物视觉筛选表型。为了说明这些方法，我们诱变蠕虫和不协调（UNC），矮胖（DPY）和变压器（TRA）突变体。

Watch this Scientific Journal Video about An Introduction to Worm Lab: from Culturing Worms to Mutagenesis at JoVE.com

An Introduction to Worm Lab: from Culturing Worms to Mutagenesis

与表型的缺陷突变体的筛选是一个简单的方法识别基因功能，在一个给定的生物过程。在这篇文章中我们将介绍如何对文化自由的生活的蠕虫（如，<em&gt; Pristionchus pacificus</em&gt;）在实验室中，并显示两个不同的诱变方法，环境管理体系和TMP /紫外线。

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