Purchase Solution

To make primers and use them for site directed mutagenesis

Not what you're looking for?

Ask Custom Question

What are the criteria for "good" primers in a PCR reaction?

Describe how you would use site-directed mutagenesis to change a BamHI restriction site into an EcoRI site.

Purchase this Solution
Solution Summary

Criteria for designing PCR primers are listed. A brief description of the site directed mutagenesis needed to change a BamHI restriction site into an EcoRI site is shown.

Solution Preview

These are the criteria for "good" primers in a PCR reaction.
i. Use a GC content of around 50%, ie don't have too many of one kind of base or your primer may bind repetitive DNA. Don't use runs of single nucleotides.
ii. Use a G or C at the 3' end to make sure base pairing is strong at the critical 3' end
iii. Use a primer length of 16nt or more, though this varies a lot with template. I usually use about 20nt. DNA has 10bp per turn so imagine the primer ...

Purchase this Solution
Free BrainMass Quizzes
Comfort Measures For Labor

Are you ready to doula someone through labor?

Light and Sight Vocabulary

This quiz introduces basic definitions of vocabulary related to light and how human eyes. This information is important for an understanding of sight.

Identifying Variables in Science Experiments, Part 2

Using sample experiments, test yourself to see if you can identify independent, dependent, and controlled variables. Identifying variables is key in understanding and developing experiments. The questions are biology related, but this can be applied to any area of science.

Do You Know Your Macromolecules?

This quiz will assess your knowledge of the macromolecules that are important to living things.

Vision and Oculomotor Control

This quiz will test the student's knowledge of the neural underpinnings of the visual system and its central pathways.

View More Free Quizzes
Related BrainMass Solutions

  • Site directed mutagenesis can be used to study ATP synthase

    This solution provides assistance determining the most likely targets of action of the particular reagent, and how one might use site-directed mutagenesis to determine whether the residue is essential for proton conduction.

  • DCCD and the F0 unit of ATP synthase

    How might you use site-directed mutagenesis to determine whether this residue is essential for proton conduction? DCCD is dicyclohexylcarbodiimide, a chemical reagent which reacts with acidic amino acid residues.

  • Fructose bisphosphate aldolase

    On that page, you will see that site-directed mutagenesis experiments were conducted on the E. coli enzyme wherein the invariant lysine (K236, in that case) was changed to an alanine (A236). In so doing, the enzyme was completely disabled.

  • Plasmid transfer

    them.

  • Designing primers for PCR: finding problems with primers

    These primers are far too GC rich. c). These primers are complementary to each other. Print out their sequences, cut out the primers and see what happens when you align them in an antiparallel fashion: base pairing allong the entire length!

  • Engineering a recombinant plasmid DNA to over-express protein

    you would like to use for this project.

  • Polymerase chain reaction

    By adding restriction enzyme sites Mary can then digest the PCR product (it needs to be run on a gel and gel purified to make sure you only have one product and get rid of excess nucleotides) and ligate it into a desired vector.

  • What is the Polymerase Chain Reaction?

    Annealing: In this step, primers that have been designed specifically for the DNA strand of interest, will hybridize with the sample DNA providing a starting point for the DNA polymerase to attach and begin the DNA replication process.

  • restriction enzyme fingerprinting

    You can also digest the BAC with the a restriction enzyme to get fragments that are 1-2kb in length and then subclone into a vector and use the primers to amplify them and then sequence them.

View More Related Solutions