User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/08/17

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LB media preparation/ Calculation of oligo dilutions

ABSTRACT

  • Preparation of LB media solid and liquid.
  • I made the proper calculations to dilute the oligos that we orderd last week.

  • Because my line of work is going to be focused in the assembly of parts I am doing other things while the synthesis arrive. Today I prepare 2 Lt of LB medium because the reservoirs of the team where scarce. I made one Lt of solid medium and other Lt of liquid medium with the following proportions:
Yeast extract 5 g/L
Casein Peptone 10 g/L
NaCl 10 g/L
-When the medium is solid I added 15 g/L of bacteriologic agar.
  • I put them in flasks and I autoclaved them for 1 hour.

  • The oligos arrived on Monday so we have to dilute them to the correct concentration to use them when necessarily.

I used a formula that says C0V0=CFVF where C0 is initial concentration, V0 is initial volume, CF is final concentration and VF is final volume. We want a final concentration of 5pM/μL and a final volume of 500μL. So by knowing the initial concentration we can use this formula to find the volume of oligo and water that has to be mixed to obtain the desired concentration.


For example:


Oligonucleotide 5158 HydEF1.Int.Rev

 -C0= 341.02 pM/mL
 -Clearing for V0 we obtain that he have to mix 7 μL of oligo and 493 of water.
  • I made a series of calculations for other oligos:
Name Reference number Oligo H2O mQ
HydEF1_Fwd 5157 17μL 483μL
TerBack_PFR_Rev 5156 15μL 485μL
TerBack_PFR_Fwd 5155 23μL 477μL
HydG.Rev 5162 18μL 482μL
HydG.Fwd 5156 16μL 484μL