Describe the ingredients necessary for a PCR reaction (such as the one in lab 8), and...

Describe the ingredients necessary for a PCR reaction (such as the one in lab 8), and why they are necessary.

Expert Solution

Template DNA , DNA polymerase, primers, deoxy nucleoside triphosphate, buffer and Mg2+ cofactor and buffer are the 6 main components of PCR.

The template strands contain our target sequence which will be amplified . The genome can be from any source, a plasmid DNA, human genome , bacterial, viral etc. Always an optimum amount is better to increase specificity . Primers are oligonucleotide sequence which binds to flank regions of target DNA sequence. It is primer which decide the nature of target sequence. Primer is designed depending on our sequence weather to amplify virulant genes, drug resistant gene etc. They are 15-30 bp long

DNA polymerase is the enzyme which is required throughout.Taq polymerase is the enzyme used which withstand in high temprature. They provide thermaltability and add nucleotide at a temprature at 70°c.dNTPs are building blocks of new DNA strand . They are of 4 nucleotides added optimal base in operation. Excess incorporation may leads to inhibition or misincorperation of bases. Magnesium is essential cofactor for function of DNA polymerase which will enable dNTPs binding. It also help in formation of phosphodiester bond . It also facilitate complex between primers and template by stabilizing negative charge . Suitable chemical environment required of polymerase chain reaction is provided by buffer. Potassium ions of KCL helps in primer annealing. Sometimes pottasium is replaced by ammonium ions which will increase specificity by destabilizing effect. It helps in maintaining ionic imbalnce and helps in amplification.

Thank you.

gladiator answered 2 years ago

Describe the ingredients necessary for a PCR reaction, and why they are necessary.

Describe polymerase chain reaction (PCR). How did we utilize this technique in lab?

As a reminder, PCR is the technique used to replicate DNA in the lab. PCR is...

As a reminder, PCR is the technique used to replicate DNA in the lab. PCR is based on the way DNA is replicated in cells. During PCR template DNA plus other ingredients are placed in a test tube and the solution goes through repeated cycles of heating and cooling. The 'ingredients' of a basic PCR reaction are: DNA template, primer, nucleotides, DNA polymerase, and any salts required for polymerase function (buffer). No ligase is required in PCR but it is...

Sudz Beer buys the necessary ingredients to brew and bottle one batch of beer for a...

Sudz Beer buys the necessary ingredients to brew and bottle one batch of beer for a total of $24,300. Each batch yields 75,960 bottles of beer. The cost per bottle is $0.32. They sell their beer to a distributor with a 35% markup, and then the distributor sells it to the retailer for a 41% markup. Because the retailer has a virtual monopoly on consumer sales, they get away with selling the beer at a 62% margin. What is the...

Lane 2 of a PCR shows no bands, indicating an unsuccessful PCR reaction. The reaction also...

Lane 2 of a PCR shows no bands, indicating an unsuccessful PCR reaction. The reaction also shows poorly separated bands (larger bands) in the reference ladder lane. Describe whether or not there is a connection between the success of a PCR reaction and how well separated the bands are in your reference ladder.

in pcr practical, Briefly describe the reaction mechanism leading to the synthesis of a new DNA...

in pcr practical, Briefly describe the reaction mechanism leading to the synthesis of a new DNA strand by Taq DNA polymerase. In your answer include: i. The mechanism of the synthesis reaction, ii. The direction of the synthesis reaction iii. The direction in which sequence information is read from the template strand.

Explain how PCR (polymerase chain reaction) works. After explaining PCR, compare PCR to the DNA replication...

Explain how PCR (polymerase chain reaction) works. After explaining PCR, compare PCR to the DNA replication that occurs naturally in living cells, making sure to give at least 4 similarities and 4 differences.

what does the polymerase chain reaction (PCR) accomplish? 1) PCR converts RNA to DNA 2) PCR...

what does the polymerase chain reaction (PCR) accomplish? 1) PCR converts RNA to DNA 2) PCR corrects mutation in a gene 3) PCR makes many copies of DNA segment 4) PCR joins short segments of DNA to make one long segment how does the ribosome select the next amino acid to add to a growing polypeptide chain? 1) the ribosome selects the amino acid that can form hydrogen bonds with the mRNA 2) the ribosome selects a tRNA that base-pairs...

prepartion of biodiesel lab Describe the role of potassium hydroxide in this reaction?

Why does DNA sequencing reaction use one primer and PCR use two primers?

Question