Cheung Lab, Day 4

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Arabidopsis prepped for dipping with Agrobacterium

On Day 4, our final day of the first week, we finished up learning about transformation methods.  We had a four day week because yesterday was July 4th, and the lab was closed to celebrate the holiday!

We started out the day by transferring our leaf-discs from co-cultivation medium with Agrobacterium to SIM-which is used to grow shoots.  The Agrobacterium were washed off of the leaf discs, using B5- which is basic growth medium.  We completed 3 washes to basically dilute the bacterium to a very low level.  The discs will stay in this medium for about a week.

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Plates in the 22 degree Celsius growth chamber

We moved our plates to the growth chamber in the basement of Lederle Graduate Research Tower.  The plates will grow in the chamber for a while.

We then started a new process of transformation, this time using Arabidopsis.  The method we used was transformation by dipping.  Basically, you take the unopened buds, and dip them in a solution of agrobacterium.  This solution will change some of the seeds.  You then screen the next generation, find the modified seeds, and grow them into plants, which produce a new generation of modified seeds.

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Mixing Agrobacterium on an orbit shaker.