SB209

Just a question, Faris and I were considering an alternative to purifying and isolating bacteriophages, do we have access to an Ultracentrifuge which reaches 13000G? We would be able to pellet bacteriophages. I am new so, I do not know about all of our lab equipment.

Today was just a bit of reading and getting some shizz together. Trying out one more method and now got a new member on team fazza so should be good. You all know Erik. We have found a protocol to extract DNA out of these phages which will be the next step then Amplifying with PCR and analysing them on a gel. This will usually take a whole day to do and is a long process and cant afford to not prepare.

So today I woke up hopeful but to only find a couple of plaques! Really thought this method was the best one also but oh well aye that’s science! At least Im capable of getting a few isolated now the upcoming weeks is going to be involved with extracting DNA out of these phages and analysing them. Data should be appearing in the near future!

Not sure if anybody is interested in this. Not sure if its real science or media exaggeration? take a look for yourselves http://www.bbc.co.uk/news/health-22967727

Hello! I think most of you know me already but incase you don’t… I’m Rachael, (just finished) 2nd year biomed.

Steve (Wade) and I are working with/for Alan and Driton on a project involving impressively fluorescent bogeys! In slightly more scientific terms we’re investigating targeted drug delivery. Alan kindly performed the geeky part of creating the GFP-fusion (GFP-green fluorescent protein) which we transformed our tame E. coli with. This has proved very successful and although not at all required SDS-PAGE has confirmed the presence of this protein in our cells so, as Alan’s said, we’re now on to up-scaling and producing masses of bright bogeys ready for some spectroscopy and purification.

On the cell culture side of things with Driton, we received two flasks of CACO-2 intestinal epithelium cells last Thursday? (I think). These are growing well and one flask has been passaged already and the other should be ready tomorrow. It’ll take approx. 3 weeks for the cells to be ready but once they are we can start performing our ‘drug trials’ to basically see if cell uptake of the protein is successful and where it goes 🙂

I’ll try and keep you updated and might even try being a little more sciencey with it!… doubt it though….

I once dissected a pig

On the other project Beth is growing plenty of cells so that we have enough material to start purifying proteins from Paracoccus denitrificans.

So, in true academic style I’ve mainly been telling other people what to do then going to meetings. But, the construct I made to express a GFP-fusion protein works (bright green cells!) so onto large scale expressions and purification. All of the credit for actually doing the work goes to Steve and Rachael though!

Alright so got some results today and I think I got around one plaque today :(. This is good in the area of method development. My second attempt has been the most successful with potentially 15 plaques! So obviously I think I have made too many tweeks to my method development at once for the third attempt so now im doing a variation from my second attempt with only one change so hopefully the results this Friday should be promising. Otherwise than that Ive been able to look at a couple papers about the PCR and running the gel section so the method development for that should be interesting and prep for that is being taken place as we speak so hopefully I’ll have some handsome looking data I can share in the near future….

So after a day of almost continuous Gram Staneing I feal that I am (finaly) semi competent at performing this test, Hooray! I am however frustrated by streak plastering ability! Whenever I perform this test The result is generally a few isolated colonies predominated by large lines of indefinable colonies that cannot be isolated. Is the result as you can imagine does not look good and is a pain to deal with. Is there anything that I can do to deal with this frustrating issue?

Today for me was really just another preparation day for making plaques! So just making my soft agar, nutrient agar and my broths. Ive also made 25 nutrient broths using the bench autoclave which was pretty cool! And on the sneaky I removed an isolated plaque made last week and placed it in a nutrient broth and incubated it for overnight so maybe I can analyse it tomorrow! If I can. Anyways night.

So, cat brush 1 completed and 1/3 way through the second. After my weekend away at my friend’s house I now have two dog brushes to analyse too. Generally I’m finding white cotton, probably my lab coat as the samples match, or could be from people wearing white cotton t-shirts now it’s summer (meant to be!) but I’ll see when I get the %fibre frequencies calculated.

Free food and alcohol!!! Now I have your attention it would be really good to see you all at the Postgraduate Symposium on Friday 12th July at Riseholme Conference Centre. There will be a range of talks and posters from postgrads in the School. There is even a prize for best student-asked question so you can be in profit for the day. And free lunch and wine. If you want to come can you please drop me an email so I can get some idea of numbers for catering etc….