commonly used solutions (2) ------ 1 ) 2 ) 3 )

 

Julie B. Wolf, UMBC

 

10 M Ammonium Acetate

To prepare a 10 M solution in 100 ml, dissolve 77 g of ammonium acetate in 70 ml of H2O at room temperature. To prepare a 5 M solution in 100 ml, dissolve 38.5 g in 70 ml of H2O. Adjust the volume to 100 ml with H2O. Sterilize the solution by passing it through a 0.22um filter. Store the solution in tightly sealed bottles at 4C or at room temperature. Ammonium acetate decomposes in hot H2O and solutions containing it should not be autoclaved.

Ampicillin

Prepare a stock of 100 mg/ml in water. Sterilize by filtration. Store at -20C but avoid repeated freeze/thaw cycles. Use at a final concentration of 100 ug/ml.

Cresol Red Loading Dye - 2.5X - for PCR reactions

1M sucrose, 0.02% cresol. Prepare 1% cresol red in water (0.5 g/50 ml). To prepare loading dye, dissolve 17 g sucrose in a total volume of 49 ml of water. Add 1 ml 1% cresol red.

EB buffer (Qiagen recipe)

10 mM Tris-Cl pH 8.3

EDTA stock

To prepare 1 liter, 0.5M EDTA pH 8.0: Add 186.1 g of disodium EDTA-2H 2O to 800 ml of H 2O. Stir vigorously on a magnetic stirrer. Adjust the pH to 8.0 with NaOH (approx. 20 g of NaOH pellets). Dispense into aliquots and sterilize by autoclaving. The disodium salt of EDTA will not go into solution until the pH of the solution is adjusted to approx. 8.0 by the addition of NaOH. For tetrasodium EDTA, use 226.1 g of EDTA and adjust pH with HCl.

6x gel loading buffer

0.25% Bromophenol blue

0.25%Xylene cyanol FF

15% Ficoll Type 4000

120 mM EDTA

IPTG

IPTG is isopropylthio-b-D-galactoside. Make a 20% (w/v, 0.8 M) solution of IPTG by dissolving 2 g of IPTG in 8 ml of distilled H 2O. Adjust the volume of the solution to 10 ml with H 2O and sterilize by passing it through a 0.22um disposable filter. Dispense the solution into 1 ml aliquots and store them at -20 ?C.

LB Medium

To make 1 liter, use 10 g tryptone, 5 g yeast extract, 10 g NaCl. Adjust pH to 7.0. Sterilize by autoclaving.

LB Agar

Dispense 15 g per liter of agar directly into final vessel. Prepare LB medium as above and add to agar. NOTE: Agar will not go into solution until it is autoclaved (or boiled). If adding antibiotics, autoclave medium first and allow to cool until warm to the touch, then add the antibiotic. Dispense about 30 ml per plate. Allow plates to dry either at 37C overnight or 20 minutes in a laminar flow hood (lids removed). Store in original Petri plate bags, inverted, at 4C for up to 2 weeks.

NaCl

To prepare 1 liter of a 5 M solution: Dissolve 292 g of NaCl in 800 ml of H 2O. Adjust the volume to 1 liter with H 2O. Dispense into aliquots and sterilize by autoclaving. Store the NaCl solution at room temperature.

NaOH

The preparation of 10 N NaOH involves a highly exothermic reaction, which can cause breakage of glass containers. Prepare this solution with extreme care in plastic beakers. To 800 ml of H2O, slowly add 400g of NaOH pellets, stirring continuously. As an added precaution, place the beaker on ice. When the pellets have dissolved completely, adjust the volume to 1 liter with H2O. Store the solution in a plastic container at room temperature. Sterilization is not necessary.

20X SB (electrophoresis buffer)

(Buffer diluted to 1X should be 10 mM Sodium hydroxide and pH 8.5 )

for 1 liter, weigh out 8 g NaOH and ~40 g boric acid - add water, dissolve and add additional boric acid until pH = 8.0; bring final volume to 1 liter.

 

 

Commonly used solutions (1)---  1 ) 2 ) 3 ) 4 ) 5 ) 6 )

 

Commonly used solutions (2)--- 1 ) 2 ) 3 )

 

Solutions  for molecular cloning---1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )