Do you need a buffer for a PCR reaction?

Do you need a buffer for a PCR reaction?

PCR buffer is necessary to create optimal conditions for activity of Taq DNA polymerase. Buffers often contain Tris-Hcl, KCl, and sometimes MgCl2.

Does PCR use a buffer?

PCR is carried out in a buffer that provides a suitable chemical environment for activity of DNA polymerase. The buffer pH is usually between 8.0 and 9.5 and is often stabilized by Tris-HCl.

What does PCR buffer contains?

The major components of PCR buffer include Tris-HCl, potassium chloride (KCl) and magnesium chloride (MgCl2). Tris-HCl and KCl are responsible for maintaining a stable pH during PCR. Magnesium ions act as cofactors for DNA polymerase so as to ensure proper DNA synthesis function of the polymerase during PCR.

What is the most important function of the PCR buffer?

PCR buffers ensure that the PCR reaction is conducted under optimal conditions. The major components of PCR buffer include Tris-HCl, potassium chloride (KCl) and magnesium chloride (MgCl2). Tris-HCl and KCl are responsible for maintaining a stable pH during PCR.

Why magnesium is used in PCR?

Magnesium is required as a co-factor for thermostable DNA polymerase. Excessive magnesium concentrations also stabilize double stranded DNA and prevent complete denaturation of the DNA during PCR reducing the product yield.

Which is the best reaction buffer for PCR?

Standard reaction buffer for PCR using FastStart Taq DNA Polymerase. Use this buffer together with FastStart Taq DNA Polymerase. For applications refer to FastStart Taq DNA Polymerase, 5 kU, GMP Grade. Unspecific endonucleases (λDNA): Not detectable in up to 20 μL after 16 hours incubation at +37°C.

Which is the 10x MgCl2 buffer for PCR?

12161516103: Will be supplied as ”PCR buffer (10X) w MgCl2”. Unit of measure is ”mL”. Standard reaction buffer for PCR using FastStart Taq DNA Polymerase.

Can a PCR reagent be autoclaved without affecting performance?

Autoclave all reagents and solutions that can be autoclaved without affecting their performance. Of course, primers, dNTPs, and Taq DNA Polymerase should not be autoclaved. Have your own set of PCR reagents and solutions that are used only for PCR. Store these reagents in small aliquots.

How to prepare PCR reagents and template DNA?

Change gloves frequently, especially if you suspect they have become contaminated with solutions containing template DNA. Always use new and/or sterilized glassware, plasticware, and pipettes to prepare PCR reagents and template DNA. Autoclave all reagents and solutions that can be autoclaved without affecting their performance.