<html>
<head>
<meta http-equiv="Content-Type" content="text/html; charset=Windows-1252">
</head>
<body dir="auto">
<div>Hi Peter: could you post the pCS2-specific "T7" sequence, please?</div>
<div><br>
</div>
<div>Thanks,</div>
<div>Dan<br>
<span style="background-color: rgba(255, 255, 255, 0);">--<br>
Daniel C. Weinstein</span>
<div><span style="background-color: rgba(255, 255, 255, 0);">Associate Professor<br>
Biology Department<br>
Queens College, CUNY<br>
<a href="x-apple-data-detectors://0/0" x-apple-data-detectors="true" x-apple-data-detectors-type="address" x-apple-data-detectors-result="0/0">65-30 Kissena Boulevard</a><br>
Queens, NY 11367-1597<br>
<a href="tel:718-997-4552" x-apple-data-detectors="true" x-apple-data-detectors-type="telephone" x-apple-data-detectors-result="0/1">718-997-4552</a> (office) /4258 (lab)<br>
Fax: <a href="tel:718-559-6745" x-apple-data-detectors="true" x-apple-data-detectors-type="telephone" x-apple-data-detectors-result="0/2">718-559-6745</a><br>
<a href="mailto:daniel.weinstein@qc.cuny.edu">daniel.weinstein@qc.cuny.edu</a></span></div>
</div>
<div><br>
On Jun 11, 2014, at 2:56 PM, "Peter Klein" <<a href="mailto:pklein@mail.med.upenn.edu">pklein@mail.med.upenn.edu</a>> wrote:<br>
<br>
</div>
<blockquote type="cite">
<div><span>Hi Jill, </span><br>
<span></span><br>
<span>I agree with Sally. But the “T7" sequence in pCS2 is not the standard T7 primer sequence, so sequencing facilities may not be able to sequence from that end with their stock primers. I think you need to provide the sequencing facility with a pCS2 appropriate
“T7" primer or use T3 (a little farther from the 3’ end). SP6 and T3 are fine. </span>
<br>
<span></span><br>
<span>I think this plasmid might be from Eddy DeRobertis lab, so you can also check with them about it.</span><br>
<span></span><br>
<span>Peter</span><br>
<span></span><br>
<span></span><br>
<span>On Jun 11, 2014, at 2:02 PM, Sally Moody <<a href="mailto:samoody@gwu.edu">samoody@gwu.edu</a>> wrote:</span><br>
<span></span><br>
<blockquote type="cite"><span>Hi Jill,</span><br>
</blockquote>
<blockquote type="cite"><span>We don't have that sequence, but in our experience it is better to send it out for sequencing rather than rely on somebody's map because things are not always what they seem. Since it is in pCS, you can use M13 primers or better
yet SP6 and T7 primers for that plasmid.</span><br>
</blockquote>
<blockquote type="cite"><span>Good luck, Sally</span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span>On Wed, Jun 11, 2014 at 1:46 PM, Jill Johnston <<a href="mailto:johnstoj@ucalgary.ca">johnstoj@ucalgary.ca</a>> wrote:</span><br>
</blockquote>
<blockquote type="cite"><span>Hello,</span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span>We have a construct called pCS-GAP43-GFP here and have no information</span><br>
</blockquote>
<blockquote type="cite"><span>about it. Does anyone have the sequence that could be sent to us?</span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span>Thanks</span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span>Jillian Johnston</span><br>
</blockquote>
<blockquote type="cite"><span>Research Associate</span><br>
</blockquote>
<blockquote type="cite"><span>Dr. Sarah McFarlane lab</span><br>
</blockquote>
<blockquote type="cite"><span>University of Calgary</span><br>
</blockquote>
<blockquote type="cite"><span>Faculty of Medicine</span><br>
</blockquote>
<blockquote type="cite"><span>Dept of Cell Biology and Anatomy</span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span>_______________________________________________</span><br>
</blockquote>
<blockquote type="cite"><span>Xenopus mailing list</span><br>
</blockquote>
<blockquote type="cite"><span><a href="mailto:Xenopus@lists.mbl.edu">Xenopus@lists.mbl.edu</a></span><br>
</blockquote>
<blockquote type="cite"><span><a href="https://lists.mbl.edu/mailman/listinfo/xenopus">https://lists.mbl.edu/mailman/listinfo/xenopus</a></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span></span><br>
</blockquote>
<blockquote type="cite"><span>-- </span><br>
</blockquote>
<blockquote type="cite"><span>Sally A. Moody, Ph.D.</span><br>
</blockquote>
<blockquote type="cite"><span>Professor of Anatomy and Regenerative Biology</span><br>
</blockquote>
<blockquote type="cite"><span>George Washington University</span><br>
</blockquote>
<blockquote type="cite"><span>School of Medicine and Health Sciences</span><br>
</blockquote>
<blockquote type="cite"><span>2300 I Street, NW, Washington, DC, 20037, USA</span><br>
</blockquote>
<blockquote type="cite"><span>202-994-2878</span><br>
</blockquote>
<blockquote type="cite"><span>Editor-in-Chief: genesis, The Journal of Genetics and Development</span><br>
</blockquote>
<blockquote type="cite"><span>Editor: Principles of Developmental Genetics, 2nd edition</span><br>
</blockquote>
<blockquote type="cite"><span>_______________________________________________</span><br>
</blockquote>
<blockquote type="cite"><span>Xenopus mailing list</span><br>
</blockquote>
<blockquote type="cite"><span><a href="mailto:Xenopus@lists.mbl.edu">Xenopus@lists.mbl.edu</a></span><br>
</blockquote>
<blockquote type="cite"><span><a href="https://lists.mbl.edu/mailman/listinfo/xenopus">https://lists.mbl.edu/mailman/listinfo/xenopus</a></span><br>
</blockquote>
<span></span><br>
<span></span><br>
<span>_______________________________________________</span><br>
<span>Xenopus mailing list</span><br>
<span><a href="mailto:Xenopus@lists.mbl.edu">Xenopus@lists.mbl.edu</a></span><br>
<span><a href="https://lists.mbl.edu/mailman/listinfo/xenopus">https://lists.mbl.edu/mailman/listinfo/xenopus</a></span><br>
</div>
</blockquote>
</body>
</html>