Soft agar assays were performed in 6-well plates. The base layer of each well consisted of 2 ml (with a final concentration of 1x) medium and 0.6% low melting point agarose (Duchefa Biochemie, Netherland). Plates were chilled at 4 °C until solid. Next, 2 ml of growth agar layer was poured, consisting of 1 × 104 cells suspended in 1x medium and 0.3% low-melting point agarose; plates were again chilled at 4 °C until the growth layer congealed. Further 1 ml of 1x medium without agarose was added on top of the growth layer. Cells were incubated at 37 °C with 5% CO2 for approximately 14–21 days, and total number of colonies were stained with 0.005% crystal violet (Sigma-Aldrich) and counted. Images were analyzed using an Olympus microscope (Olympus, Japan) and Image-Pro Plus 4.5 software (Media Cybernetics Inc., Rockville, MD, USA). Assays were repeated a total of three times.
Low melting point agarose
Manufactured by Duchefa Biochemie
Low melting point agarose is a type of agarose, a polysaccharide derived from red algae, that has a lower melting point compared to traditional agarose. It is commonly used as a gelling agent in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using low melting point agarose
1
Soft Agar Assay for Colony Formation
2
Soft Agar Colony Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soft agar assays were performed in 6-well plates. The base layer of each well consisted of 2 ml with final concentrations of 1× medium and 0.6% low-melting point agarose (Duchefa, Haarlem, The Netherlands). Plates were chilled at 4℃ until solid. Next, a 1 ml growth agar layer was poured, consisting of 5×104 cells suspended in 1× medium and 0.3% low-melting point agarose; plates were again chilled at 4℃ until the growth layer congealed. A further 1 ml of 1× medium without agarose was added on top of the growth layer. Cells cultures were maintained at 37℃ for 14 days with exchanging the media for every 3 days. Total colonies were stained with 0.005% crystal violet (Sigma, St Louis, MO, USA) and counted in five randomly fields. Images were analyzed using Image-Pro plus 4.5 software (Media Cybernetics).
3
Soft Agar Assay for Cell Transformation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soft agar assays were performed in 6-well plates containing a base layer of 2 ml (at a final concentration of 1X) medium and 0.6% low melting point agarose (Duchefa Biochemie, Netherland). Plates were chilled at 4 °C until the media solidified, and then a growth layer of 2 ml growth agar containing 1 × 104 cells suspended in 1X medium and 0.3% low-melting-point agarose was added. Plates were chilled again at 4 °C until the growth layer congealed. An additional 1 ml of 1X medium without agarose was added on top of the growth layer. Cells were incubated at 37 °C in 5% CO2 for approximately 14–21 days, and colonies were stained with 0.005% crystal violet (Sigma-Aldrich) and counted. Images were analyzed using an Olympus microscope (Olympus, Tokyo, Japan) and Image-Pro Plus 4.5 software (Media Cybernetics Inc., Rockville, MD, USA). Assays were performed in triplicate.
4
Soft Agar Assay for Cell Transformation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Soft agar assays were performed on 6-well plates. The base layer of each well consisted of 2 ml (with a final concentration of 1×) medium and 0.6% low melting point agarose (Duchefa Biochemie, Netherland). Plates were chilled at 4 °C until solid. Next, 2 ml of growth agar layer was poured, consisting of 1 × 104 cells suspended in 1× medium and 0.3% low-melting-point agarose; plates were again chilled at 4 °C until the growth layer congealed. Further 1 ml of 1× medium without agarose was added on top of the growth layer. Cells were incubated at 37 °C with 5% CO2 for approximately 14–21 days, and a total number of colonies were stained with 0.005% crystal violet (Sigma-Aldrich) and counted. Images were analyzed using an Olympus microscope (Olympus, Tokyo, Japan) and Image-Pro Plus 4.5 software (Media Cybernetics Inc., Rockville, MD, USA). Assays have been repeated a total of three times.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!