What kind of errors would be produced by excess taq in PCR?

In summary, adding excess taq pol to a PCR reaction can result in errors due to the inhibitory effect of glycerol, which is present in the enzyme's storage solution. It is recommended to never add more than 10% of enzyme to a reaction to avoid this issue.
  • #1
Wek
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What kind of errors would be produced by "excess" taq in PCR?

Not sure if this belong in the Homework section. Put it here since it's not homework.

What kind of errors will I get by adding 6.75µL taq pol to +50µL of cDNA? The master mix will be added to the cDNA resulting in an excess of taq pol. Only 2 and 4µL of the taq+cDNA mixture will be used for each reaction not the entire +50µL.
 
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  • #2


There is not information here to answer your question. As a rule you never add more than 10% of enzyme to your reaction, this is because your enzyme is stored in glycerol and that this is inhibitory to the reaction.
 

What kind of errors would be produced by excess taq in PCR?

If there is an excess of taq in PCR, it can lead to several types of errors, including:

1. How does excess taq affect the specificity of PCR?

When there is too much taq in the PCR reaction, it can increase the likelihood of non-specific binding, resulting in false amplification of non-target DNA fragments.

2. Can excess taq cause PCR artifacts?

Yes, excess taq can lead to the formation of primer dimers and other PCR artifacts, which can interfere with the accurate amplification of the target DNA sequence.

3. What impact does excess taq have on the sensitivity of PCR?

An excess of taq can decrease the sensitivity of PCR by using up the available primers and other reagents, making it difficult to detect the target DNA in the sample.

4. Does excess taq affect the efficiency of PCR?

Having too much taq can actually decrease the efficiency of PCR by competing with other components in the reaction, resulting in a less efficient amplification process.

5. How can excess taq be avoided in PCR?

To prevent excess taq in PCR, it is important to carefully measure and optimize the amount of taq used in the reaction, following recommended guidelines and protocols.

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