Subcloning by restriction digestion is a common laboratory technique and Creative Enzymes will discuss how to transfer cDNA from one plasmid to another. However, the same technique can be used to move promoters, selection markers or any other DNA components between plasmids. Creative Enzymes provides you with a variety of plasmid cloning service by restriction enzymes service to meet your scientific research needs.

Figure 1.Recombinant DNA technology.

Sepcific Plasmid Cloning Service by Restriction Enzymes Approaches We Offer

We first perform a DNA analysis that allows you to identify which restriction sites are present in a given sequence. When choosing a restriction endonuclease, you will want to select one that has

• Flank your insert, but do not cut within your insert
• Cuts at the desired location in your recipient plasmid (usually at the polyclonal site (MCS)), but not elsewhere in the plasmid
• Will result in your insert being in the correct orientation in the recipient plasmid.
• Conform frame to the tag or fusion protein in the recipient plasmid

If you cannot find an enzyme that meets these criteria, we can provide the following services.

• Adding the desired restriction site to the flank of the insert

We use PCR-based cloning and add the restriction site to the end of the oligonucleotide.

• Adding the desired restriction site to your recipient plasmid

We use annealed oligonucleotide clones to modify the MCS of your recipient plasmid.

Workflow

• Digesting your DNA.

Set up restriction digests for your donor and recipient plasmids. Because some DNA is lost during the gel purification step, it is important to digest a large amount of starting material.

• Separate insert fragments and vectors by gel purification.

Run the digested DNA on an agarose gel and perform gel purification to isolate the DNA.

• Ligate your insert into your vector.

Perform DNA ligation to fuse the insert to the recipient plasmid.

• Transformation.

Transform your ligation reaction into the bacterial strain of your choice

• Isolation of the completed plasmid.

Finally, you need to pick individual bacterial colonies and check that they are successfully ligated.

Service Process

Our Features

• We have accumulated many years of essence in the field of plasmid cloning service by restriction enzymes service, helping customers to speed up project development and improve the overall success rate of the project.
• At Creative Enzymes, simply let us know your specific project needs. We will suggest the best strategy for you.

Deliverables

• Related results raw data
• Data analysis
• Image and result analysis
• Results analysis
• Activity assessment report
• Summary of relevant parameters

Why Choose Us?

Creative Enzymes is a company that provides professional and comprehensive plasmid cloning service by restriction enzymes service. We have years of experience to meet your specific project needs in using enzyme research to add value to your research projects. Creative Enzymes can provide you with personalized solutions to help you thrive every step of the way around your interest in your workflow. If you would like to know more about this service, please feel free to contact us.