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<html><body style="word-wrap: break-word; -webkit-nbsp-mode: space; -webkit-line-break: after-white-space; "> Hi Jonathan,<div><br></div><div>Those concentrations are reasonable, assuming that you are injecting about 1 nl/embryo. If you read any of the original papers on the Tol2 transposon you will find details. For instance, look at our Tol2kit paper:</div><div><br></div><div><span style="font-size:12.0pt;font-family:"Times New Roman"; mso-ansi-language:EN-US;mso-fareast-language:EN-US">KM Kwan et al.</span><span style="font-size:12.0pt;font-family:"Times New Roman"; mso-ansi-language:EN-US;mso-fareast-language:EN-US"> (2007) The Tol2kit: a multisite Gateway-based construction kit for <i>Tol2</i></span><span style="font-size:12.0pt;font-family:"Times New Roman";mso-ansi-language:EN-US; mso-fareast-language:EN-US"> transposon transgenesis constructs. <b><i>Developmental Dynamics</i></b></span><span style="font-size:12.0pt;font-family:"Times New Roman"; mso-ansi-language:EN-US;mso-fareast-language:EN-US"><i>, </i></span><span style="font-size:12.0pt;font-family:"Times New Roman";mso-ansi-language:EN-US; mso-fareast-language:EN-US">236:3088-99.</span> </div><div><br></div><div> Or, read the Tol2kit wiki:</div><div><br></div><div><a href="http://chien.neuro.utah.edu/Tol2kitwiki/">http://chien.neuro.utah.edu/Tol2kitwiki/</a></div><div><br></div><div>--Chi-Bin Chien</div><div><br><div><div>On Jan 27, 2009, at 7:56 AM, Jonathan Schumacher wrote:</div><br class="Apple-interchange-newline"><blockquote type="cite"><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Greetings,</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; min-height: 14px; "><br></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">I am a rookie with zebrafish transgenics and have been given a project</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">to inject embryos with a construct in conjunction with transposase</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">cRNA.<span class="Apple-converted-space"> </span>I don't see in the literature where the concentration of</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">construct & transposase are described.<span class="Apple-converted-space"> </span>I've been using 20ng/uL for</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">both construct & transposase.<span class="Apple-converted-space"> </span>Does this seem like a reasonable</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">concentration?<span class="Apple-converted-space"> </span>I saw on another posting that Dr. Lister described</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">using ng/uL concntrations.</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Thoughts?</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; min-height: 14px; "><br></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Many thanks!</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; min-height: 14px; "><br></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Jonathan Schumacher</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Research and Development Scientist</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Anatomic Pathology Molecular Diagnostics</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">ARUP Institute for Clinical & Experimental Pathology</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">500 Chipeta Way</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Salt Lake City, UT 84108</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Telephone: 801-583-2787 extension 2958</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Fax: 801-584-5109</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Email: <a href="mailto:schumaj@aruplab.com">schumaj@aruplab.com</a></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; min-height: 14px; "><br></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">_______________________________________________</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; ">Zbrafish mailing list</div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; "><a href="mailto:Zbrafish@net.bio.net">Zbrafish@net.bio.net</a></div><div style="margin-top: 0px; margin-right: 0px; margin-bottom: 0px; margin-left: 0px; "><a href="http://www.bio.net/biomail/listinfo/zbrafish">http://www.bio.net/biomail/listinfo/zbrafish</a></div> </blockquote></div><br></div></body></html>

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