Diluting Ampicillin into LB AgarLPS Flow Cytometry
Posted 20 May 2011 - 02:47 PM
I'm new to this website and I found it helpful in reading some of the interested topics
just to mention that I started my undergrad research in purifying RNA mainly H69,
I really need some help in some of the calculations concerning O.D readings
So basically I ran a 1 mL reaction for 4-5 hours, centrifuge to remove any salts, do ethanol precipitation and disolve the pellet in dH2O.
then I take the O.D of the sample and I read it as let say it's 100 O.D at 260nm witha volume of 70 uL
now if I want to make the reading to be 20 OD (since i need to run it in a small 9 by 7 gel) how much of the 100 O.D sample should be taking and how much should i dilute it to.
If there is a website that you think would be helpfull to me to look at, please post it
I will be posting more question, cz I really need help with this as an undergrad
Posted 25 May 2011 - 01:27 AM
Its best to make a new post with questions like this.
Posted 10 December 2011 - 07:00 PM
Posted 12 February 2012 - 09:56 PM
I am kinda confused with the dilutions.....
1.I need to make a 0.02% of Bromophenol blue (working solution) in a 5Xor 10X buffer. I made 5X buffer and then made a 2% firstly.
by adding 0.16 gm to 6.4ml 5X buffer (made 8ml).
Now I need to make a 0.02 or0.01 final dilution. How do i do this?
2. I know the final concentration of my RNA extracted. I need to know how much I am loading or using /ul. I get so annoyed calculating in terms of ug/ul.
Can someone help me please.
Posted 14 February 2012 - 08:23 AM
concentration, in your case, is weight per volume. g/l=mg/ml=ug/ul. if you know the concentration then you know how much you are loading/ul.
genius does what it must
i do what i get paid to do
Posted 20 February 2012 - 05:01 PM
Thanks for ur explanation. I kept myself cool and worked out....I could get the concentration and the loading amount too.
Posted 25 September 2012 - 03:56 PM
I have a 5mg/mL LPS concentration in 20 ul
I have 200ul of DC cells in my well (2 X 10^5 cells total in each well)
I want to make a working stock of 50ug/mL LPS and transfer 10ul into my wells.
From there I want to make 1:5 dilutions of LPS (50, 10, 2, 0.4)
How do I start to make my LPS stock?
Please explain in detail as I have the calculation but I want to figure this out completely.
Please read the rest of this thread - it will answer your questions with a bit of thought. Bob
Posted 28 September 2012 - 08:55 AM
I'm very new to the lab, but I'm starting to learn alot.
Just doing some routine cloning, I'm fine with everything except after weeks, I still haven't figured out how to dilute my ampicillin stocks! This is both embarrassing and ridiculous so I'm asking for some help if anyone can teach me on hwo to do it and I can apply this to everything else with regards to dilutions. I have no problems diluting 50X TAE to 1X TAE but I don't know why with ampicillin it is such a huge problem ):
Anyway, so I have made up 100mg/ml of Ampicillin stocks from powder.
I want to make up LB agar plates using 30mls of liquified LB agar.
How much ampicillin should I add if i want a concentration of 100ul/ml?
I know my stocks is at 1000X how do I go from here? Should I duilte to make working stocks of 1 or 10X first?
I am making 10 plates so that makes it to 300mls of LB Agar but I usually aliquot the ampicillin separately into plates that have been poured out with 30mls of LB agar Already (still in liquid form)
Appreciate all help! thanks a million!
Presuming you want ug/ml not ul/ml use the 1:1000 dilution. Keep your units the same for all variables. Bob.
Posted 28 September 2012 - 08:53 PM