Would it be possible to kill bacteria without damaging their protein structure? There are several ways to kill bacteria without damaging protein structure or function. For example, consider the following frequent ones: 1. First, heat inactivation: Read More
In E. coli prokaryotic expression vector pET32a, does the TrxA tag lead to non-specific binding in WB and ELISA assays? A TrxA tag is often used to make sure that recombinant proteins fold and Read More
How do I perform a restriction digest on total DNA isolation from mammalian cells? Isolating whole DNA from mammalian cells and performing a restriction digest is a multi-step process. It is important to note that Read More
Mice Anesthetization Protocols Anesthetizing mice is a common practice in biomedical research for various purposes, including surgery, imaging studies, or the collection of tissues. Several methods are available, including injectable and inhalant anesthetics. 1. Injectable Anesthetics: Read More
SDS-PAGE Gel Preparation 1. Prepare the separation gel (10%). Mix in the following order: H2O 4.1 mL Acrylamide/bis (30% 37.5:1; Bio-Rad) 3.3 mL Tris-HCl (1.5 M, pH 8.8) 2.5 mL SDS, 10% 100 µL N,N,N′,N′-tetramethylethylene-diamine (TEMED) (Bio-Rad) 10 µL Ammonium Read More
Tamoxifen Preparation and Treatment in Mice for Genetic Recombinations Rationale: Tamoxifen is used to kick-start genetic recombinations in genetically engineered mouse models (GEMMs) and to activate genes through a cascade of events. Here is a Read More
DNA Quantification using Qubit Fluorometer The Qubit Fluorometer is a device from Thermo Fisher Scientific used to quantify DNA, RNA, and protein in samples by using a fluorescent dye that binds to these molecules. The Read More
DNA Quantification using Nanodrop spectrophotometer The nanodrop spectrophotometer is commonly used to quantify DNA concentration and assess purity. Here is a basic outline of the process: 1. Calibrate the machine: It’s usually done by using Read More
DNA Extraction Protocol in Detail Here is a widely accepted protocol for the extraction of DNA from cells, especially mammalian cells. This method is called the phenol-chloroform extraction method. Materials 1. Cell culture or tissue sample 2. Read More
RNA Extraction Protocol in Detail Let’s dive into details using a widely accepted protocol based on an RNA extraction kit (which utilizes the organic extraction method with TRIzol, which is a monophasic solution of Read More
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