What is Platinum Taq polymerase?
Platinum Taq DNA Polymerase, DNA-free, is a recombinant Taq polymerase complexed with a proprietary antibody that blocks polymerase activity at ambient temperatures and dissociates after the initial denaturation step at 94°C. Find out more at www.thermofisher.com/dna-free.
What is the role of Taq polymerase?
Taq polymerase denotes the heat-stable DNA polymerase extracted from the thermophilic bacteria Thermus aquaticus. It is used to automate the repetitive steps in the polymerase chain reaction (PCR) technique, an extremely important method of amplifying specific DNA sequences.
What is hot start Taq polymerase?
Hot Start Taq DNA Polymerase is a mixture of Taq DNA Polymerase and an aptamer-based inhibitor. The inhibitor binds reversibly to the enzyme, inhibiting polymerase activity at temperatures below 45°C, but releases the enzyme during normal cycling conditions, allowing reactions to be set up at room temperature.
What are nucleotides in PCR?
Nucleotides (dNTPs or deoxynucleotide triphosphates) – single units of the bases A, T, G, and C, which are essentially “building blocks” for new DNA strands.
What does 2X master mix mean?
2X Taq Mix is a premixed, ready-to-use solution containing Taq DNA Polymerase, dNTPs, Mg2+ and Reaction Buffer at optimal concentrations for efficient amplification of DNA. templates by PCR.
Why is Taq polymerase used in a polymerase chain reaction PCR )?
Due to its key role in synthesizing and amplifying new strands of DNA, Taq DNA Polymerase is essential to Polymerase Chain Reaction (PCR). Like other DNA polymerases, Taq Polymerase can only produce DNA if it has a primer, a short sequence of 20 nucleotides that provide a starting point for DNA synthesis.
How is primer design for the gateway attB primers?
Correct design of attB primers for amplification, cloning and expression of a gene in Gateway requires consideration of the proper placement of protein expression elements (ribosome recognition sequences, start codon, stop codons, reading frame considerations etc.) with respect to the attB recombination sites.
What is the hot start property of Platinum Taq DNA polymerase?
The hot-start property of Platinum Taq DNA Polymerase allows for convenient reaction assembly at room temperature. Just as with Taq DNA Polymerase, Platinum Taq DNA Polymerase has a non-template–dependent terminal transferase activity that adds a 3′ deoxyadenosine to product ends, and has a 5′→3′ exonuclease activity.
Which is an example of an attB primer pair?
B1 B2 ATG Stop Table 1 provides examples of primer pairs (attB1 and attB2) which can be used to amplify genes for specific protein expression formats. Shine-Dalgarno (E. coli ribosome binding sequence) and Kozak (eukaryotic ribosome recognition sequence) show are typical sequences.
What are the shared sequences of att1-4?
Here is a set of “shared” sequences–the core stretch that is shared between attB/P/L/R for att1-4. These are useful for stitching together entry and destination sequences to calculate the sequence of a final expression clone. Below are all of the att sequences (attB, P, L, and R) used in the original three-part multisite Gateway system.