Sunday, 7 August 2016

Ways To Ensure Experimental Consistency #3

Commercial Kits

There are many commercial kits available to streamline and simplify various experimental procedures including DNA/RNA extraction, plasmid preparation, cloning, immunoassays, reporter assays, and so forth.

Commercially available kits are certified for quality and have proven efficacy so long as the instructions for use and handling are adhered to. However, there are a few important points to note in regards to reagent and sample preparation, points that some product manuals may not necessarily provide information on. 

* Ensure that components of buffers and reagents are in solution. With some buffers, you may see precipitates. For example, lysis buffers containing SDS tend to develop white precipitates when the room temperature is low. This is the SDS falling out of solution. In such instances, warm up the buffer in a water bath before use. 

* Ensure correct storage of kit components. Make sure that after opening and using individual kit components that they are stored at the recommended temperature. While a kit may arrive at room temperature, after a buffer is opened it may need to be stored at 4 degrees.

* Note the expiry date of the kit and/or specific components.

* If you are required to prepare a component that is not provided (e.g. addition of 3’ A overhangs to inserts for TOPO cloning) be sure to use reagents that are of a quality and purity compatible with subsequent parts of a kit. 


* Ensure that samples are extracted in a buffer that is compatible with the reagents supplied. A notable example is the Bradford Protein Assay, which is incompatible with some detergents and substances.

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