Anaida Walia's Profile

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How to Dephsophorylate DNA Using Calf Intestinal Phosphatase

When you dephosphorylate a vector for cloning, you remove the 5′ phosphate groups required by DNA ligase to join the phosphodiester DNA backbone together. This prevents your vector from ligating back to itself during the ligation step and decreases your background. Various alkaline phosphatases exist, including calf intestinal phosphatase (CIP), Shrimp and Antarctic phosphatases. The […]

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In DNA / RNA Manipulation and Analysis 12th of December, 2016
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How to Disinfect Plant Explants for Tissue Culture

Prior to using plant explants, the surface of the explant needs to be disinfected to prevent introduction of unwanted microbes into your culture. Usually, you use chemicals (called sterilant liquids) at appropriate concentrations and duration of application to disinfect the explants. A Sterilizer’s Dream: The Ideal Disinfectant The efficacy of a disinfectant depends on many […]

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In Cells and Model Organisms 20th of September, 2016
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A Quick Guide to Screening by Restriction Digestion

PCR screening is great for identifying potential positive clones after cloning/subcloning. However, you’ll still want to confirm that the DNA sequence is correct for any positives identified. The easiest and quickest way to do this is by restriction digestion. This guide will tell you about the theory and practice of using this technique. The Theory […]

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