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>20bp Insertion Mutagenesis - How to?

>20bp Insertion Mutagenesis - How to? - Biology Forum

>20bp Insertion Mutagenesis - How to? - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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Old 10-30-2012, 06:42 PM
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Default >20bp Insertion Mutagenesis - How to?



Hi All,

I need to insert 21-30 bps. I inserted 1-2 nucleotides before by simply using primers with these extra bps but 30 bps sounded a little too much to do it with primers. How should I do it? I have these in mind:

1) I can do it with long primers (e.g. 70 bps) with insertion bps in the middle.
2) I can do it sequentially with shorter primers with 5-6 insertions at a time (prefer not to)
3) I can do it with a kit.

I haven't used a kit before and don't know how they work. I m lazy and preferred to ask here instead of reading bunch of different kits' protocols. To be honest I prefer not to buy a kit also.

What is your suggestion? Do you think I can do it with 1st option?

Thanks in advance,

Engin
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Old 06-13-2013, 06:04 PM
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Default Re: >20bp Insertion Mutagenesis - How to?

In science, peptide synthesis is characterized as the formation of a peptide bond between two amino acids. While the definition of a peptide is not definitive, it usually refers to flexible (little secondary structure) chains of up to 30-50 amino acids. <a href="http://www.unitedpeptide.com">United Peptide</a> provide peptide synthesis services.

The ability to form peptide bonds to link amino acids together is over 100 years old, although the first peptides to be synthesized, including oxytocin and insulin, did not occur for another 50-60 years, demonstrating the difficult task of chemically synthesizing chains of amino acids (1). In the last 50 years, advances in protein synthesis chemistry and methods have developed to the point where peptide synthesis today is a common approach in even high-throughput biological research and product and drug development (2).

The benefit of peptide synthesis strategies today is that besides having the ability to make peptides that are found in biological specimens, creativity and imagination can be tapped to generate unique peptides to optimize a desired biological response or other result. This page highlights the important aspects of peptide synthesis, the most common methods of synthesis and purification and the strengths and limitations of the respective strategies.
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