Extraction from agarose gel of biopart J04450. Continuation of the assembly of the constructions to characterize the transcriptional terminator B0015. Inoculation of TAP medium with Chlamydomonas reinhardtii to obtain the transcripts HydEF and HydG.
Today Héctor Medina and I, helped by Enrique Paz, performed electrophoretic gel band extraction with a kit. Before using the kit, we first made gel electrophoresis to prepare the bands. The procedure was the same as any other gel I've made, but it was performed with low melting point agarose (1% gel) and it was made in the cold room at 80 volts during 2:30 hours. Here is the electrophoretic gel that contains the purified fragments as well as other stuff that belongs to Melissa Molho and Vladimir Muñoz.
Lane #1 contains 1kb DNA ladder, lane #2 contains the band extraction of the J04450 digestion that corresponds to the fragment that contains the RFP. All the other lanes contain other stuff that are not mine. As I have been already told, this procedure reduces significantly the amount of DNA that is wanted to isolate.
I left incubating at 25°C (lab's room temperature u_u) 100 ml of TAP medium [1] with ampicillin (500 μl per 100 ml of medium) with Chlamydomonas reinhardtii 137C; I wanted to measure its optical density but I was not able to homogenize the cells in the medium. I will be measuring its OD until it reaches OD750 = 1. At this point I will pass the 100 ml volume with cells to a 100 ml volume flask and let them overnight to obtain the anaerobiosis environment that Maluye recommended to do (as she does not have argon to use it as this paper protocol states [2])
UNAM Genomics Mexico 2011
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