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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16079" title="hello, Am working on expressing protein in Hep2 cells using a retroviral vector (pGCsamEN). Has anyone else worked with this vector or transfection of mamalian cells with retroviral vectors, if so would you please tell a rough protocol.Regards,Vipin..." target="_self">Retroviral transection</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16077" title="I would like to perform expression analysis from a highly polymorphic gene with 525 documented mutations and about 100 SNP\'s within a 10.3 kb region of the X chromosome. Patient studies indicate multiple SNP\'s present within 5\' gene flank, 5\'UTR and i..." target="_self">Cloning whole gene (10.3kb) in an expression plasmid vector</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16065" title="Hi All,I’m new to recombinant protein production and I am a bit confused about the transfection of a cell line. We have a CMV plasmid in the lab which has previously been used to transfect COS-7 cells using Lipofectamine. Do the cells continue to divide a..." target="_self">After Transfection</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16056" title="I was wondering if anyone has used beta-mercaptoethanol as part of their routine transformation process? It has been reported to increase transformation efficiency, but I am not sure if this is only for the particular cells I am using (XL10-gold) or if th..." target="_self">B-Me used for transformations?</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16037" title="With the Promega pGEMT-easy cloning kit, I have trouble getting positive clones. I use electroporation instead of heat shock for transforming my JM109 cells. For this, I precipitate the ligation and resuspend it in water prior to electroporation. I wonder..." target="_self">pGEMT easy TA cloning</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16031" title="Hi,I am currently trying to clone a large insert of 7.5kb into a large vector of 12kb. My current protocol is to excise the insert from pBluescript using pac1 and asc1, gel extract using qiagen kit, phenolchloroform extract and precipitate and then ligate..." target="_self">Help cloning large insert into large vector</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16026" title="Dear everyone, I have a problem about pIRES2-DsRed2 (5.3kb,clotech) double digestion. I used XhoI and BamHI to digest the pIRES2-DsRed2 and I got two bands, which are both around 2500bp. I should to be only one band. And then I extracted the fragment (5.3..." target="_self">problem about pIRES2-DsRed2 double digestion</a><br />');
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document.write('<li class="rss-item"><a class="rss-item" href="http://www.protocol-online.org/forums/index.php?showtopic=16025" title="hi.. all, can anyone help me.... i have a doubt that.. is it okey doing cloning with topo xl for the genomic DNA which has restricted with RSa1 or Alu1 . The smear concentrating near 1kb. and also what is the faint bands that is there in the smear. whethe..." target="_self">topoxl cloning</a><br />');
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