<html><body><div style="color:#000; background-color:#fff; font-family:HelveticaNeue-Light, Helvetica Neue Light, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif;font-size:16px"><div dir="ltr"><span>Dear Katherine,</span></div><div dir="ltr"><span><br></span></div><div dir="ltr" id="yui_3_16_0_1_1425198104034_48548"><span id="yui_3_16_0_1_1425198104034_48547">You are completely right that it is difficult to get enough DNA for sequencing from blood, but yes PCR amplification can help get sufficient template for sequencing (if it is only for a specific gene target). I would suggest to use buffy coat rather than whole blood for PCR. Your results will be dramatically improved.</span></div><div dir="ltr" id="yui_3_16_0_1_1425198104034_48546"><span><br></span></div><div dir="ltr" id="yui_3_16_0_1_1425198104034_48545"><span id="yui_3_16_0_1_1425198104034_48544">All the best</span></div><div></div><div id="yui_3_16_0_1_1425198104034_48535"> </div><div class="signature" id="yui_3_16_0_1_1425198104034_47833"><div id="yui_3_16_0_1_1425198104034_48536"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_48538"><em id="yui_3_16_0_1_1425198104034_48537">Prof. Sarman Singh, MBBS, MD, FAMS, FIMSA, FRSTMH, FRSC, FATP, FSIIP<br></em>Head, </span></div><div id="yui_3_16_0_1_1425198104034_48540"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_48539">Division of Clinical Microbiology & Molecular Medicine,<br>All India Institute of Medical Sciences, </span></div><div id="yui_3_16_0_1_1425198104034_47842"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_47841">New Delhi (India)<br>Editor: Journal of Laboratory Physicians</span></div><div id="yui_3_16_0_1_1425198104034_47840"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_47839">Phone: +91-11-26594977, 26588484<br>Fax: +91-11-26588663, 26588641<br></span></div><div id="yui_3_16_0_1_1425198104034_47838"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_47837">https://www.researchgate.net/profile/Prof_Sarman_Singh</span></div><div id="yui_3_16_0_1_1425198104034_47836"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_47835"><span style="font-family:Calibri;line-height:22.7199993133545px;" id="yui_3_16_0_1_1425198104034_47834">Personal Website: </span><a rel="nofollow" target="_blank" href="http://www.drsarmansingh.com/" style="line-height:22.7199993133545px;cursor:pointer;font-family:Calibri;">www.drsarmansingh.com</a><span style="font-family:Calibri;line-height:22.7199993133545px;"></span></span></div><div style="line-height:22.7199993133545px;font-family:Calibri;" id="yui_3_16_0_1_1425198104034_47832"><span style="font-size:small;color:rgb(164, 96, 22);" id="yui_3_16_0_1_1425198104034_47831">-BMJ Award 2014, Excellence in Innovative Healthcare Technology<br style="line-height:22.7199993133545px;"></span><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-MedAchiever's Merit Award 2014, Indian Medical Association</span><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-<i style="line-height:22.7199993133545px;">Chikitsa Ratan</i> Award 2014, Delhi Medical Association</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-Dr. BK Aikat Oration 2008, Indian Council of Medical Research</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-Dr. BC Roy Award 2008, Medical Council of India</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-<i style="line-height:22.7199993133545px;">Vigyan Ratan Samman</i>, 2007, UP Council of S &T</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-Dr. BP Pandey Oration Award 2006, Indian Society of Parasitology</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-Biotech Process, Product & Commercialization Award, 2006, Department of Biotechnology</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-Merit Award 1998, International Immnunocompromised Host Society USA</span></div><div style="line-height:22.7199993133545px;"><span style="font-size:small;color:rgb(164, 96, 22);">-Young Scientist Award 1996, International Immnunocompromised Host Society USA</span></div></div></div></div> <div class="qtdSeparateBR"><br><br></div><div class="yahoo_quoted" style="display: block;"> <div style="font-family: HelveticaNeue-Light, Helvetica Neue Light, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif; font-size: 16px;"> <div style="font-family: HelveticaNeue, Helvetica Neue, Helvetica, Arial, Lucida Grande, sans-serif; font-size: 16px;"> <div dir="ltr"> <font size="2" face="Arial"> On Friday, March 6, 2015 6:51 AM, "Sayler,Katherine A" <saylerk@ufl.edu> wrote:<br> </font> </div> <blockquote style="border-left: 2px solid rgb(16, 16, 255); margin-left: 5px; margin-top: 5px; padding-left: 5px;"> <br><br> <div class="y_msg_container"><div id="yiv1597281505">
<style type="text/css"><!--#yiv1597281505 P{margin-top:0;margin-bottom:0;}--></style>
<div dir="ltr">
<div>Dear readers, </div>
<div><br>
</div>
<div>Thank you sincerely for your time. I am a tick-borne disease biologist by trade, so this is outside of my realm.
<br>
</div>
<div><br>
<font size="3" face="Calibri"><span style="font-size:12pt;"><span dir="ltr"><span style="background-color:white;"></span></span></span></font></div>
<div><font size="3" face="Calibri"><span style="font-size:12pt;"><span dir="ltr"><span style="background-color:white;">I am trying to confirm some preliminary data that suggests the
<em>Leishmania</em> spp. are circulating in limited geographic locales in southern Florida. </span><font size="3"><span style="background-color:white;">Is it at all reasonable to expect to detect the DNA of the trypanosomatids in the blood of infected wildlife?
My suspicion is that this is completely improbable, as it appears that in order to detect DNA in enough quantity or quality for sequencing, you need cultured organisms. From sandflies it appears that it is possible to PCR amplify and sequence directly from
the vector, but never from the reservoir (or potential reservoir) host. Is this true? Any advice or insight would be greatly appreciated and I am tremendously naïve and this is clearly not my area of expertise</span></font></span></span></font>.<br>
</div>
<div><br>
</div>
<div>Thank you again for reading my message,</div>
<div>Katherine<br>
</div>
<div id="yiv1597281505Signature">
<div style="font-family:Tahoma;font-size:13px;">
<div style="font-family:Tahoma;font-size:13px;">
<div>
<div style="font-family:Tahoma;font-size:13px;"><br>
</div>
<div><font size="2" face="Arial">Katherine Sayler, M.Ed., Ph.D.</font></div>
<div><font size="2" face="Arial">Postdoctoral Associate | Department of Wildlife Ecology and Conservation</font></div>
<div><font size="2" face="Arial">Molecular Ecology Laboratory | 2322 Mowry Rd. Gainesville, FL 32611</font></div>
<div><font size="2" face="Arial">Phone: 352-871-3259 </font></div>
</div>
</div>
</div>
</div>
<br><br>--<br>
<i> This email was sent by icb.usp.br </i>
</div>
</div><br>_______________________________________________<br>Leish-l mailing list<br><a ymailto="mailto:Leish-l@lineu.icb.usp.br" href="mailto:Leish-l@lineu.icb.usp.br">Leish-l@lineu.icb.usp.br</a><br><a href="http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l" target="_blank">http://lineu.icb.usp.br/cgi-bin/mailman/listinfo/leish-l</a><br><br>--<br>This email was sent by icb.usp.br<br><br><br></div> </blockquote> </div> </div> </div> </div></body></html>