prabhubct

I always make up and sterilise more than I need (plain LB) and store it at room temp for sometimes up to months at a time.

I usually make between 200 and 300ml in a 500ml schott bottle, then microwave it to melt it before pouring. I'll often then not use the entire 300ml and so that will reset and I will remelt it again later when I need it. In this case, I have on occasion got a contaminant on the top of the set agar but this is rare (like maybe twice in 6 years?).

I've never had difficultly redissolving the agar, you just have to make sure that you heat it up to near boiling and mix by swirling really well. I hold it up to the light to make sure the texture is consistent, you can easily see when it is ready. It would take me maybe 5 minutes to do.

I prefer this to making plates and storing them in the fridge long term. Especially as I don't use many plates at a time and use a range of different antibiotics depending on what I'm doing.

That, and the condensation completely grosses me out!! I have issues with that kind of thing, gives me the heebie jeebies!

Not that I am really an expert on this but I've had my share of frustrating moments especially at congresses feeling like a kindergarden child compared to the annoyingly witty professors with huge departments and annoyingly perfect publications. I think every language in the world has a version of the saying: "those that know do, those that don't know teach". In other words once you get your hands dirty, you will know exactly what you need to know. You will also know very well which papers to read and what to remember.
My 2c. Best of luck.

ok, this one's for ex-perlmunky....a bit late but who knows...he might just pay us a surprise visit...

this here's a mangling of K'naan's song 'Hurt me Tomorrow':


Leave Us Tomorrow (casey's version)

I used to think that PERL is just embedded in a trinket

            and Linux is Charlie's bestfriend with the blanket

C++ for me is but an almost F

and banks with no money but only genes so what the F?

Yo Fasta Blast aint as scary as Gangsta Blast

No idea 'bout Python Programming, Unix, source-coding,

what's white hat hacking .......plus Java is the coffee I drink in the morning

but now I'm a bit enlightened......

If what we read is true

Then we know what you came to do

You may be tired but we're hoping that you

would stick around and may be consider....

Ref: This ain't a good time

But when is it ever

I know the perfect time

And dude that's never

So don't you dare leave us now

Throw your posts on the ground

Cause tonight ain't the night for sorrow

But you can leave us tomorrow....

Good question. I just asked my colleague now, he said he often uses Salmonella in electroporation and he prepares them himself. They work well, and he says you can make them chemically competent too. They must work.

"There is no teacher but the enemy. No one but the enemy will tell you what the enemy is going to do. No one but the enemy will ever teach you how to destroy and conquer. Only the enemy shows you where you are weak. Only the enemy tells you where he is strong. "


Mazer Rackham, O Card's Ender's Game

The Monster: For as long as I can remember people have hated me. They looked at my face and my body and they ran away in horror. In my loneliness I decided that if I could not inspire love, which is my deepest hope, I would instead cause fear. I live because this poor half-crazed genius, has given me life. He alone held an image of me as something beautiful and then, when it would have been easy enough to stay out of danger, he used his own body as a guinea pig to give me a calmer brain and a somewhat more sophisticated way of expressing myself.

Young Frankenstein

"It's best to have failure happen early in life. It wakes up the Phoenix bird in you so you rise from the ashes."
Anne Baxter

Transfection, sorry.

The limiting size is not the problem here, the question is not whether it's possible to transfect with two plasmids, the question is, if he can transfect two separate plasmids, one with obviously different efficiency AND use the second plasmid to calculate the efficiency of the first.

It would be like measuring how much salt can dissolve in an amount of water by measuring how much sugar dissolves there.

depends on how large the protein is.

you can reduce the acrylamide concentration of the gel

add up to 0.05% sds and 20% methanol to the transfer buffer

transfer for more time (how much more should be determined by trial)

remember that you may never get 100% transfer

here is a useful book:

Depends what you want to do with them. I am doing online courses without getting a degree:

https://www.coursera.org/courses

I only do them to specialize/learn new stuff. They do not offer a degree or certificates (most courses don't) but the knowledge is the same as if you would take the course at Princeton or Stanford etc. In the end of the day, to do properly a job you do not need a framed paper on a wall, but knowledge.

Hello.
I would like to thank everybody for expressing his/her sympathy to my case. Special thanks for the practical clues too.
Well, teaching sounds great but in my country for 4th-5th year the only one University that provides study of Natural sciences has plenty of unoccupied positions for students - nobody wants to study life sciences, chemistry or physics because they are difficult and have no realisation here. Plus, all teaching positions are occupied until the rest of the eternity.
Research.....ah, the life here is as expensive as in the rest of EU (for some things like energy we pay much more) but salaries....I would be lucky if I get 250 Eu per month. And the system for grants is still very corrupted. And the whole society here is against people like me - here only the practical jobs are respected. This is why I wrote my topic....and I think I got some clues :-)
Finally, I am 35 years old and it is absolutely neccessary to give a birth as soon as possible.
In the name of God, I have been working all my life for this!
You see - it is not a matter of psychological depressia but of a real problem......that I hope to resolve somehow :-)
One more time thank all of you :-)

My supervisor just asked me to help her to write a recommendation letter for one of her students and I found this PDF file and wanted to share it with you. I found the sample letters so well written:

http://www.hhmi.org/...oads/letter.pdf

biofuel, on Jun 13 2009, 04:46 PM, said:

Having spent 18 years in industry and 13 years in academia...these in my opinion are the pro's and con's

Industry: Pro's
Permanent position.
Training infra-structure
Money and resources
Team atmosphere
Higher Salaries
Better pensions/health cover/SAYE share schemes

Cons:
No academic freedom
Very specialised area of screening/research
Difficulty in getting a position in the first place
Possibility of takeover/merger/relocation........redundancy.


Academia: Pro's
Academic freedom
A more relaxed environment

Con's
Tenured positions/short term contracts
Poor pay structure
Very little career structure
Competition for grant money
Pressure to publish and in good journals
Normally not working in a multi disciplinary environment.

You make your own judgements...but in my opinion go for the industry jobs first.

Kindest regards

Rhombus

And here an adapted version of an old song from Alice Cooper: School's Out

Well we're not choosy
All the nerds and geeks
bein' all that nosey
'Cause they found no sheiks
Well they can't fund ya
Can't find work
If that don't suit ya
Try to smirk

Lab's out for summer
Lab's out forever
Lab's been blown to pieces

No more pipets
No more papers
No more student’s dirty coats

Well we got no class
And we got no results
And we got no publication
We can't even think of a word that rhymes

Lab's out for summer
Lab's out forever
Lab's been blown to pieces

No more enzymes
No more digests
No more technician's dirty coats

Out for summer
Out till fall
We might not go back at all

Lab's out forever
Lab's out for summer
Lab's out with fever
Lab's out completely

The next time you prepare your LB plates take one and keep it Overnight at 37°C. If something grows on it discard the rest. This will save you time and money.