Biomedical Engineering Reference
In-Depth Information
Hydrogen
Oxygen
Nitrogen
Carbon
Phosphorus
T
A
C
G
Pyrimidines
Purines
Figure 1.1
(See companion CD for color igure.) . Double-stranded. DNA. (B-form).. Reprinted. under. terms. of. creative.
commons..attribution-share.alike.3.0.unported.license;.image.created.by.Richard.Wheeler..(From.Wheeler,.Image.
ile.DNA.Structure.+.Key.+.Labelled.pn.NoBB.png,.created.by.Richard.Wheeler.(Zephyris).and.used.under.terms.
of. Creative. Commons. license,. 2010,. http://en.wikipedia.org/wiki/File:DNA_Structure%2BKey%2BLabelled.
pn_NoBB.png,.accessed.July.23,.2012.)
or.volume.in.deionized.water..The.staple.strands.in.DNA.origami.are.typically.<50.bp.in.
length;. because. these. strands. are. typically. used. in. 20-100-fold. excess,. the. purity. of. the.
crude.desalted.oligos.is.suficient.to.give.high.yields.of.properly.assembled.origami..For.
DNA. origami,. a. synthesis. scale. of. 100.nmol. for. each. of. the. staple. strands. will. provide.
enough.material.for.over.50.separate.annealing.runs,.and.each.annealing.run.will.make.
over.1.mL.of.2.nM.origami,.enough.material.for.numerous.AFM.imaging.experiments..At.
2011.prices,.the.cost.of.staple.strands.for.a.typical.(90.×.60.nm).origami.at.100.nmol.scale.is.
about.$1800.
The.array.of.techniques.used.to.handle.and.characterize.DNA.are.beyond.the.scope.of.
this. chapter;. a. good. sourcebook. of. methods. for. handling. DNA. oligos. is. the. Cold Spring
Harbor Molecular Cloning .handbook.(Sambrook.and.Russell.2001)..Chemically,.DNA.is.quite.
sturdy.(Green.et.al..2010);.it.can.survive.aeons.of.time.or.be.boiled.for.hours.(Miller.1990)..
However,. DNA. is. vulnerable. to. metal-catalyzed. hydrolysis. and. light-. and. free-radical-
induced.cleavage.reactions,.and.it.is.tasty.to.bacteria,.which.secrete.endo-.and.exonucle-
ases.that.will.cleave.the.sugar-phosphate.backbone..ss.DNA.is.more.easily.damaged.than.
double-stranded.DNA..ss.oligos.can.be.preserved.for.years.by.(a).storage.in.a.laboratory.
freezer.away.from.light;.(b).autoclaving.laboratory.equipment.such.as.pipet.tips.that.will.
touch.the.oligos,.particularly.for.preparation.of.stock.solutions.that.must.be.stored.longer.
than.a.day.or.so;.and.(c).sterilization.of.buffers.(autoclave.or.ilter.through.0.45.μm.syringe.
ilter,. store. in. fridge).. Working. buffers. should. include. a. small. concentration. (∼1.mM). of.
ethylenediaminetetraacetic.acid.(EDTA).to.chelate.undesired.metal.ions.
 
Search WWH ::




Custom Search