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<div>Thx!<br>
<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>
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On Jun 11, 2014, at 3:18 PM, "Takuya Nakayama" <<a href="mailto:tn8t@virginia.edu">tn8t@virginia.edu</a>> wrote:<br>
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<div><span>That's what I was about to point out too.</span><br>
<span>In case you wonder, here is the "T7" primer which we've been using for </span><br>
<span>pCS2+ sequencing: 5'-AATACGACTCACTATAG-3' (just shorter version of </span><br>
<span>original T7).</span><br>
<span></span><br>
<span>Takuya</span><br>
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<span>On Jun 11, 2014, at 2:49 PM, Peter Klein wrote:</span><br>
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<blockquote type="cite"><span>Hi Jill,</span><br>
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<blockquote type="cite"><span>I agree with Sally. But the “T7" sequence in pCS2 is not the </span><br>
</blockquote>
<blockquote type="cite"><span>standard T7 primer sequence, so sequencing facilities may not be </span><br>
</blockquote>
<blockquote type="cite"><span>able to sequence from that end with their stock primers. I think you </span><br>
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<blockquote type="cite"><span>need to provide the sequencing facility with a pCS2 appropriate “T7" </span><br>
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<blockquote type="cite"><span>primer or use T3 (a little farther from the 3’ end). SP6 and T3 are </span><br>
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<blockquote type="cite"><span>fine.</span><br>
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<blockquote type="cite"><span>I think this plasmid might be from Eddy DeRobertis lab, so you can </span><br>
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<blockquote type="cite"><span>also check with them about it.</span><br>
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<blockquote type="cite"><span>Peter</span><br>
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<blockquote type="cite"><span>On Jun 11, 2014, at 2:02 PM, Sally Moody <<a href="mailto:samoody@gwu.edu">samoody@gwu.edu</a>> wrote:</span><br>
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<blockquote type="cite"><span>Hi Jill,</span><br>
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<blockquote type="cite"><span>We don't have that sequence, but in our experience it is better to </span><br>
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<blockquote type="cite"><span>send it out for sequencing rather than rely on somebody's map </span><br>
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<blockquote type="cite"><span>because things are not always what they seem. Since it is in pCS, </span><br>
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<blockquote type="cite"><span>you can use M13 primers or better yet SP6 and T7 primers for that </span><br>
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<blockquote type="cite"><span>plasmid.</span><br>
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<blockquote type="cite"><span>Good luck, Sally</span><br>
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<blockquote type="cite"><span>On Wed, Jun 11, 2014 at 1:46 PM, Jill Johnston </span><br>
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<blockquote type="cite"><span><<a href="mailto:johnstoj@ucalgary.ca">johnstoj@ucalgary.ca</a>> wrote:</span><br>
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<blockquote type="cite"><span>Hello,</span><br>
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<blockquote type="cite"><span></span><br>
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<blockquote type="cite"><span>We have a construct called pCS-GAP43-GFP here and have no information</span><br>
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<blockquote type="cite">
<blockquote type="cite"><span>about it. Does anyone have the sequence that could be sent to us?</span><br>
</blockquote>
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<blockquote type="cite"><span></span><br>
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<blockquote type="cite"><span>Thanks</span><br>
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<blockquote type="cite"><span>Jillian Johnston</span><br>
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<blockquote type="cite"><span>Research Associate</span><br>
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<blockquote type="cite"><span>Dr. Sarah McFarlane lab</span><br>
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<blockquote type="cite"><span>University of Calgary</span><br>
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<blockquote type="cite"><span>Faculty of Medicine</span><br>
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<blockquote type="cite"><span>Dept of Cell Biology and Anatomy</span><br>
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<blockquote type="cite"><span>_______________________________________________</span><br>
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<blockquote type="cite"><span>Xenopus mailing list</span><br>
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<blockquote type="cite"><span><a href="mailto:Xenopus@lists.mbl.edu">Xenopus@lists.mbl.edu</a></span><br>
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<blockquote type="cite"><span><a href="https://lists.mbl.edu/mailman/listinfo/xenopus">https://lists.mbl.edu/mailman/listinfo/xenopus</a></span><br>
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<blockquote type="cite"><span>-- </span><br>
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<blockquote type="cite"><span>Sally A. Moody, Ph.D.</span><br>
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<blockquote type="cite"><span>Professor of Anatomy and Regenerative Biology</span><br>
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<blockquote type="cite"><span>George Washington University</span><br>
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<blockquote type="cite"><span>School of Medicine and Health Sciences</span><br>
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<blockquote type="cite"><span>2300 I Street, NW, Washington, DC, 20037, USA</span><br>
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<blockquote type="cite"><span>202-994-2878</span><br>
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<blockquote type="cite"><span>Editor-in-Chief: genesis, The Journal of Genetics and Development</span><br>
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<blockquote type="cite">
<blockquote type="cite"><span>Editor: Principles of Developmental Genetics, 2nd edition</span><br>
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<blockquote type="cite"><span>_______________________________________________</span><br>
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<blockquote type="cite"><span>Xenopus mailing list</span><br>
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