New Project

Hello again,

Well I wanted to start my blog with good news, that this week I started a new project beside my bioluminescent bacteria project, which I’m going to explain what it is about.

But before talking about my new project, let me tell you what I did related to bioluminescent bacteria project. This week I did the transformation bacteria experiment again but this time only for Agrobacteria, and Josh told us to change the few steps so maybe the new way gives us some result because so far we didn’t get any result from the old experiment. So what I did different this time was after I placed the mixtures of transformation solution and plasmid DNA (in +pGLO tube only) on ice, then rapidly transferred them to a hot water bath (95 ºC) and this time instead of 50 seconds I kept them there for 10 minutes, and then placed them back on ice again . After two minutes, that I incubated them on ice. I did the same steps as before and added 250µl of LB Nutrient Broth. After 10 minutes, I started adding the solution from tubes to agar plates and spread it across the plate with a sterile loop. After 24 hours in the 37 ºC incubator, I checked the Agrobacteria plates with UV light but unfortunately they didn’t glow!

Agrobacteria plates

And now what’s my second project?

Matt and Josh gave me a new project “Allelopathy test” which means I’m going to spending a little bit in the Phoenix College garden to do the Allelopathy test. But before Matt giving me any further information about this project and how to do the experiment, I needed to know what is “Allelopathy”? So after what he explained to me and do some research I find out that, “Allelopathy” refers to the beneficial or harmful effects of one plant on another plant, both crop and weed species, by the release of chemicals from plant parts by leaching, root exudation, volatilization, residue decomposition and other processes in both natural and agricultural systems (ufl.edu). Which means that different plant parts include leaves, flowers, root, and also soil can have allelopathic activity and that can affect many aspects of plant ecology.

According to Matt the reason that I’m doing this project is that it seems there’s a toxin in PC garden that inhibit seed germination and seedling growth. So what I suppose to do it’s to do a leaf extraction from 4 different trees that we have in PC garden ( Sycamore, Sun flower, Orange, and Oak), then prepare 100 petri dishes with radish, lettuce, chard and carrot seeds, 5 plates of each with 25seeds on them. After that I’ll water the seeds with extract of each tree, and then I have to wait for the germination. So far, I cut all my filter paper and labeled all the petri dishes with seeds on them. Hopefully next week I’ll start the leaf extraction.

100 petridishes with radish, lettuce, chard and carrot seeds

Second week of school, first week of internship

I’m glad to be back and I’m really happy that I started my internship since this week, which is the second week of spring semester (hopefully my last semester), so that gives us more time to learn from Josh and Matt and do more cool experiments. I’m so excited about our project this semester; I’ll keep my fingers crossed to get the results from bioluminescent bacteria because so far we couldn’t make it glow!

So I started my first week with making Tryptic Soy Broth (TSB). Tryptic Soy Broth (TSB) is a nutritious medium that supports the growth of a wide variety of microorganisms, especially common aerobic and facultatively anaerobic bacteria (bd.com). The steps were pretty much the same as TSA that I did before - weighting 30.0 g of the powder and add it into the 2L flask, containing 1L distilled water. Then you heat it up and warm slightly to completely dissolve the powder. 

Tryptic Soy Broth 

Heat up the solution 

Before autoclaving it, I made the solution to three racks of test tubes with autoclave tape on top of them. Finally after 46 minutes autoclaving, the TSB’s test tubes were ready. I left them for a couple minutes at room temperature and then put them in incubator.  

TSB test tubes ready for autoclave

TSB test tubes after autoclave

But besides working on bioluminescent bacteria and TSB in the first week, my sister and I checked the cadavers and sprayed the cadaver with moistening solution. We also tried to do the blood test ourselves to find out our blood type. According to the results, I am blood type A+.

I also did transformation bacteria for E.Coli and Agrobacteria two times during last week. Normally bacteria exchange genetic material with other bacteria and they try to change to the one that can survive better. Transformation in bacterial cells occurs when the cell joins naked DNA into its genetic material. I placed the mixtures of transformation solution and plasmid DNA (in +pGLO tube only) on ice, then rapidly transferred them to a hot water bath (42ºC ) for about fifty seconds, and then placed them back on ice again – this procedure is called heat shock and “increases the permeability of the cell membrane to DNA”. After two minutes, I incubated them on ice. I added 250µl of LB Nutrient Broth. After 10 minutes, I started adding the solution from tubes to agar plates and spread it across the plate with a sterile loop. After 24 hours in the 37 ºC incubator, I checked the plates with UV light and the +pGLO for E.Coli was glowing.