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A number of assay techniques rely on the need for divalent metal ions to promote the activity of polymerases. Isothermal amplification techniques which are conducted under isothermal conditions in the absence of polymerases in the reaction mixture, have the potential to become even more attractive with the development of such techniques. Herein we report a method to enable isothermal amplification of nucleic acids to be accomplished by thermal cycling in the presence of potassium chloride (KCl) alone, thus avoiding the need for magnesium ions. Kinetic analysis of the amplification data determined the reaction parameters for a ttDNA repeat primer set, and these were used to develop isothermal amplification protocols using the ‘hot hot hot’ method, with and without KCl, with two different enzymes, leading to a total of two isothermal amplification protocols. The ‘hot hot hot’ protocol with Taq polymerase was an effective amplification system, and using this protocol Taq polymerase could be replaced with a variety of enzymes.Q:
Why do I have to add explicit forward declarations to this project in VS2012?
I have the following files that I am trying to compile in VS2012. The first file generates a compilation error that I am not clear on, yet the second does not:
PoEPDL.h:
#ifndef PoEPDL_h
#define PoEPDL_h
class Solvent;
class Phase;
class BoundaryConditions;
class PhaseStack;
class KCl;
class Methane;
class Water;
class PoEPDL
{
public:
PoEPDL(int numLayers=1);
PoEPDL(const PoEPDL& PDL);
~PoEPDL();
void solve();
private:
double *PDL; // Pass your own PDL data structure
};
#endif
PoEPDL.cpp:
#include “stdafx.h”
#include “PoEPDL.h”
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