Tabaluga

hobglobin, on 04 May 2013 - 12:12 PM, said:

nope....but you're close...

Should work fine, just watch out for compatibility of the secondaries.

You can include polls when starting a new topic or add polls into an existing topic. I don't know if that is what you desired.

Also, when we upgrade the forum program next time, we will have a new function which allows users to vote for the best answer chosen from all replies to a question, something similar to Yahoo answers.

pellets

plastics? QED...you and your pretty weird and cheap taste...

The pink/red/yellow colour is most likely a pH indicator called phenol red.  This is pinkish purple at high pH, red at about pH 7 and yellow at low pH.  The colour change is dependent to some extent on the amount of CO2 in the incubator  as dissolved CO2 forms carbonic acid, which along with the bicarbonate in the medium causes a buffering to about pH 7 with 5-10% CO2 (depends on the exact amount of bicarb in the medium).  A low CO2 reading could easily give you a purple colour - the same thing happens when you leave the medium open on the bench as there is about 0.1% CO2 in normal air.  Also note that the humidity of the air in the incubator can affect the CO2 reading from the incubator - make sure the humidifier in the bottom has some water in it.

The first thing you should do is culture some of the cells without antibiotics (if your sterile technique is any good you shouldn't need them for culturing at all) and see if you get a contaminant growing - these will often cause medium colour changes, especially towards basic pH's.  The presence of antibiotics in the medium can suppress but not eliminate low level bacterial contaminants, which can still have an effect on the cells.

Dear Guys,
I know this is out of ur discussion, but regarding converting RPM into G or vice verse, I just measure the radius of my centrifugal ring, not the whole machine, just the rotor radius with a ruler or something, put the data on RPM-G converter like these website:
http://www.endmemo.com/bio/grpm.php
http://www.geneinfin...p/sp_rotor.html

then I got the required number in G or in RPM

student guide for PCR optimization-

http://www.babec.org..._Guide_2012.pdf

and PCR calculator-

http://www.sigmaaldr.../pcr-tools.html

Dear metaltemujin,

To answer your question " how to improve your bio skills"

There are many to learn. The most important to start with is basic lab techniques. These may include:-

Basic pipetting skills
Serial Dilutions
Western Blotting Basic cell and tissue culture
Methods of sterilisation
Basic microbiology
Basic molecular biology

.......the list is endless


Then you have to learn about setting up experiments:-

Positive and negative controls
Optimisation steps
The critical path for your experiments
Time management skills
Hypothesis lead experiments
Accuracy as well as precision....i.e. your experiments must be reproducible

......the list is endless

Then you have to learn to present your research/results to your collegues....and then further afield:-

Statistical analysis
Presentation skills
What content to put in AND LEAVE OUT
Anticipating ackward questions
Looking for weaknesses in the results
Further experiments that need to be done

....the list is endless

Then if you are lucky you can contribute or write papers to go into high impsct journals:-

Again most of the above will come into that.

The most important thing I can impress on young people coming into science is the importance of which research group they join. I was very lucky in my career that the first lab head I worked for won a Nobel prize for physiology in the 1980's. This set the standard for the whole department and the post doc I worked for spent many yeras with me teaching me the basics as listed above. You can read many books and scientific papers but the most important thing is

EXPERIENCE....EXPERIENCE.....EXPERIENCE

I hope some of this is useful

Kindest regards

Uncle Rhombus

over-read this, sorry. Other common problems with these description are overloading the wells and protein contaminations, but the latter would be surprising with a PCR sample.
Sharper bands you can get e.g. if you start with 80 V until the loading dye is in the agarose and then increasing to 120 V (and if TAE buffer I wouldn't increase more), fast loading of all the wells (avoiding diffusion), a sufficiently long polymerisation time of the gel. And running the gel not too long, as then diffusion becomes a factor.

Very unlikely but, are you you looking at a temporally expressed gene with multiple isoforms.  With the template DNA you are using would a different isoform be expressed at this time point that is slightly shorter?  How are you generating your template DNA?  Maybe your target gene is not fully being reverse transcribed?  

1-RT-PCR primers are usually designed to have a product of 100-150Kb.  Quadruple check your gene from Pubmed with your primers by ClustalW (just to ensure there binding site).

2- Not necessarily.  Depending on the sequence of your primers, it may just have a high affinity for your gene of interest.  

*If all else fails, just redesign your primers.  I hope this helps.

Listen type into google search as follows: Cytokine filetype:pdf  

The part filetype will give you just PDF files and gives you a smile in your face:). You can do this also with doc, xls, etc. Enjoy the discovery.


Books:
Cytokine Protocols (Methods in Molecular Biology)

http://elib.fk.uwks....ogy Series].pdf

http://www.ctf.edu.t...1_Cytokines.pdf