I would bring up the DNA in TM buffer (10 mM Tris, 5 mM MgCl2) or similar and anneal under dilute conditions to favor hairpin formation over dsDNA.
Fast cooling will also favor hairpin formation, so you may try heating to 95° and then cooling on ice, or using a short gradient on a thermocycler. You can check your work on native PAGE (including appropriate controls, like oligos annealed under dsDNA-favoring conditions) On Thu, Mar 17, 2011 at 4:23 AM, dengzq1987 <[email protected]> wrote: > Dear all, > > Recently,i purchase Oligonucleotide from company.the sequence is > TTTTTTTTTTGCGTAC GCAC GTACGC .i want to perform self-annealing process to > form the following second structure . > 5' TTTTTTTTTTGCGTACGC > ||| ||| ] > 3' CGCATGCA > > i hope that most of the Oligonucleotide can form this structure,how can i > achieve this? any adivice is appreciated. > > > > best wishes. >
