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Why are two different primers required for the polymerase chain reaction?
Must use the forward and reverse primers to bind to complementary sequence at the 3' end of the template strand - each NEW strand is built in 5' to 3' direction. They flank the targeted gene region - must attach one to each strand of the target DNA.
What are the five necessary components of DNA synthesis reaction?
DNA polymerases, RNA primers, deoxyribonucleoside 5'-triphosphates (or dNTPs), Mg2+ ions, and a DNA template strand.
Does PCR require knowledge of the DNA sequences at the ends of the region to be amplified?
Yes, the primers need to anneal at the correct sites on the template strand for the specific region to be amplified. For the primers to attach to a specific site, they need to be in the correct sequence -- one that is opposite to the template sequence.
What is replicating the lagging strand of DNA that is adding bases in the 3-5 direction utilizes what?
The lagging strand of DNA is replicated using a process called Okazaki fragments. These are short DNA fragments synthesized in the 5' to 3' direction by DNA polymerase, and are subsequently joined together by DNA ligase to form a continuous strand.
What are okazaki fragments used to elongate?
Okazaki fragments are used to elongate the lagging strand. These fragments are used as primers for RNA polymerase to fill up the gaps in the newly formed complimentary DNA on the lagging strand. DNA ligase then seals up the gaps.
Why are two different primers required for the polymerase chain reaction?
Must use the forward and reverse primers to bind to complementary sequence at the 3' end of the template strand - each NEW strand is built in 5' to 3' direction. They flank the targeted gene region - must attach one to each strand of the target DNA.
Which DNA polymerase is responsible for removing primers and replacing them with DNA on the lagging strand during DNA replication?
The DNA polymerase responsible for removing primers and replacing them with DNA on the lagging strand during DNA replication is called DNA polymerase I.
Why are leading and lagging strand primers removed instead of being joined with Okazaki fragments during DNA replication?
Leading and lagging strand primers are removed during DNA replication because they are only needed temporarily to initiate the synthesis of new DNA strands. Once the Okazaki fragments are synthesized, the primers are no longer necessary and must be removed to allow for the joining of the fragments into a continuous DNA strand.
Do primers anneal to newly synthesized strands?
Yes, primers anneal to the newly synthesized DNA strands during the process of polymerase chain reaction (PCR). Primers provide the starting point for DNA polymerase to initiate synthesis of the new DNA strand.
What sequence in human DNA is the forward primer complementary to?
forward primers are complementary to anti sense strand of the dsDNA
What is the correct sequence of events that occur in a PCR reaction?
In a PCR reaction, the correct sequence of events is denaturation, annealing, and extension. Denaturation involves heating the DNA to separate the strands. Annealing involves cooling the reaction so primers can bind to the DNA. Extension involves DNA polymerase synthesizing a new strand of DNA using the primers as templates.
Why RNA primers are needed during DNA replication and what is their fate?
DNA polymerase cannot begin the synthesis of new DNA.To synthesis a new strand of DNA ,RNA primer is required.The complementary RNA nucleotides,that are added opposite to the single strand of parent DNA are the RNA primer.
What are the five necessary components of DNA synthesis reaction?
DNA polymerases, RNA primers, deoxyribonucleoside 5'-triphosphates (or dNTPs), Mg2+ ions, and a DNA template strand.
What is universal primer?
Universal primers are really not 'universal' in the sense that they will bind to anything. Universal is kind of a misnomer. Really, universal primers are PCR/sequencing primers that bind to a sequence found in many plasmid cloning vectors, most of which are derived from pUC vectors (which in turn come from pBR322). These sequences were defined as good PCR and sequencing sites as they flank the multiple cloning site where an inserted DNA sequence would be put. You can now buy these universal primers from various companies. You can see that these primers are called universal because they can be used to amplify or sequence any insert that is put in the multiple cloning site.
Why do you need to add primers DNA polymerase and nucleotides to your PCR mixture?
NEED OF PRIMER IN PCR-It is because the polymerase enzyme we use in the PCR only extend a DNA strand but not initiate its synthesis. So, to initiate the synthesis of DNA strand onto a template strand we require primers.
Does PCR require knowledge of the DNA sequences at the ends of the region to be amplified?
Yes, the primers need to anneal at the correct sites on the template strand for the specific region to be amplified. For the primers to attach to a specific site, they need to be in the correct sequence -- one that is opposite to the template sequence.
Role of additives in PCR?
Reactants: (dNTPs, template DNA (to be amplified), primers(bind to DNA to begin elongation of strand), DNA Polymerase (elongate DNA), & MgCl2) in buffer + H2O
