title: Eurogentec S. pombe ORF array (K130C)
status: Public on Aug 11 2005
contact.city: Stockholm
contact.first: Karl
contact.last: Ekwall
contact.email: karl.ekwall@ki.se
contact.address: Alfred Nobels Alle 7
contact.country: Sweden
contact.institute: Karolinska Inst
contact.phone: +46 8 6089133
organism: Schizosaccharomyces pombe
description: 11520 ORFs were spotted in duplicate on glass slide
manufacture_protocol: Primary PCR: The PCR reactions are performed using tagged genome specific primers in the presence of genomic DNA (provided by your laboratory). We normally achieve a success rate of 93 % of PCR reactions. For the failed PCR reactions we perform three different protocols of PCR conditions, which include different MgCl2 concentrations and different Taq polymerase. We should normally get an additional 2 % working and reach a success rate of 95 % in total. We check 5 % of the total oligos by Mass Spectrometry and will re-synthesize maximum 2 % of the oligos based on a newly designed primer. We will perform the PCR reactions with the new designed oligonucleotides with the standard PCR protocol. Following this process we reach usually 97 % success rate.