Our Engineer-it Kits provide you with all of the necessary components so that you can learn how to make the “houses” that lab bacteria grow in (LB agar petri dishes), incubate and grow the bacteria, make the bacteria able to take up DNA (make the chemically competent), and transform the bacteria with DNA (put a DNA plasmid with an extra trait into the cells). What is the end goal?
At the end of the Engineer-it Kit journey you have some LB agar plates that have little spots of bacteria on them. These are called bacterial colonies and are a result of a single bacteria that you engineered with the plasmid, multiplying into millions to form a small mound on the plate.
We’ve designed the Engineer-it kits so that we can help you gauge how expert you have become! A simple way to do this is to count the number of colonies that you got:
Zero: Try again! Every scientist, bioengineer, researcher, has tried a transformation and had cells not grow! Some suggestions:
-always use blank cells that have been growing for ~16 hours. If you’ve grown cells for more than 24 hours, it won’t work very well.
-when making competent cells, the solution should be cloudy with cells. If it is not cloudy, collect some more cells and mix them into the transformation buffer
-when making competent cells, you should not see large clumps of cells floating around. If you do, vigorously mix them some more!
-always keep the samples cold — try to keep your transformation buffer on the cold station while making your competent cells
One to Five: Excellent! You successfully engineered cells! In order to see the trait fully develop, you may need to incubate them for 48–72 hours. If you have engineered the cells with a colour pigment, after 48 hours you can turn off the incubator and leave the bacteria to grow at room temperature. This can help the pigment creation and make the colour POP.
Six to Twenty: Wow! You’re getting pretty good. This probably isn’t your first time, and it shows you that practice pays off! Did you try experimenting with the protocol? Perhaps you make competent cells with blank cells that grew only 14 hours??? How else can you get a higher transformation efficiency??
More than Twenty: You’re reaching professional level with your technique! Have you tried different plasmids? Did you know that larger plasmids are more difficult to put in bacteria than small ones?