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Soft Agar Assay Protocol. 1. Preparation of Base Agar: a. Dissolve 1% agarose (Difco Agar Noble) in sterile H20, cool to 42°C in water bath. Use autoclaved 125 5 Jul 2012 This protocol provides a general workflow for establishing a soft-agar colony formation assay. Materials and Reagents. Agarose-LE I'm using LMP Agarose (AgarPlaque Plus Agarose, BD Bioscience) for the assay. Lower layer: 0.5 I'm trying to overcome few problems with the soft Agar assay: . www.protocol-online.org/cgi-bin/prot/view_cache.cgi? 9 Sep 2015 I am having a bit of trouble counting colonies on my soft agar assay. The base is made from 1% agar and the top 0.6% agarose, I topped up media . I will give your protocol for staining a go also, I think the destaining will be 15 Jun 1998 Soft Agar Assay for Colony Formation. Protocol. Introduction In vitro cellular (e.g. agar or agarose), however when these cell lines are Prepare 1% noble agar by adding 1 g of noble agar to 100 ml of deionized water. NOTE: Noble agar will not dissolve completely with Agarose is frequently used instead of agar. 2. Cells are mixed in ~0.33% agarose/agar and layered on top of the base layer of 0.5% (or higher) agarose/agar. 3. 12 Aug 2014 Soft Agar Protocol. Materials for colony formation in soft agar assay Combine 0.8 g agarose with 25 ml of ddH2O in small glass bottle Soft agar assay protocol. Things to prepare: 2xDMEM; 1.8% agarose; ddH2O autoclaved. Procedure: Make bottom agar (0.6%):. Melt the 1.8% agarose 27 Apr 2015 Hi, I did soft agar assay on MCF-7 cells. I used 0.7% and 0.5% for plate 1 and for plate 2 0.5 and 0.35% of agarose for bottom and upper layers respectively. 5000 There is a detail protocol of soft agar assay on JOVE
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