BISC219/F12: Media Recipes

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(New page: {{Template:BISC219/F12}} <div style="padding: 10px; width: 720px; border: 5px solid #7E354D;"> == Ingredients and Stock Concentrations for some of the reagents used in Series 3 == {| bord...)
(Ingredients and Stock Concentrations for some of the reagents used in Series 3)
Line 7: Line 7:
! Reagent !! Composition   
! Reagent !! Composition   
|-
|-
-
! Solution 1*   
+
! Miniprep Solution 1*   
| 50 mM glucose<br> 10 mM EDTA (Ethylenediaminetetraacetic acid) <br> 25 mM Tris, pH 8   
| 50 mM glucose<br> 10 mM EDTA (Ethylenediaminetetraacetic acid) <br> 25 mM Tris, pH 8   
|-
|-
-
! Solution 2*
+
! Miniprep Solution 2*
| 0.2 M NaOH <br> 1% SDS (sodium dodecyl sulfate)
| 0.2 M NaOH <br> 1% SDS (sodium dodecyl sulfate)
|-
|-
-
! Solution 3*
+
! Miniprep Solution 3*
| 2.7M Potassium Acetate <br> 6.6M Acetic Acid
| 2.7M Potassium Acetate <br> 6.6M Acetic Acid
|-
|-
Line 21: Line 21:
! Nematode Growth Medium Lite(NGM lite)- Available in powder form from US Biological Cat. #N1005 (http://www.usbio.net)
! Nematode Growth Medium Lite(NGM lite)- Available in powder form from US Biological Cat. #N1005 (http://www.usbio.net)
| 0.3% NaCl<br> 1.7% Bacto-agar <br> 0.25% Bacto-peptone <br> 0.1% (vol/vol)cholesterol
| 0.3% NaCl<br> 1.7% Bacto-agar <br> 0.25% Bacto-peptone <br> 0.1% (vol/vol)cholesterol
 +
! Chemotaxis media
 +
| 2% bactoagar <br> 5 mM potassium phosphate buffer, pH = 6.0 <br> 1 mM CaCl<sub>2</sub> <br> 1 mM MgSO<sub>4</sub>
|-
|-
! Ampicillin*
! Ampicillin*

Revision as of 11:40, 10 October 2012

Ingredients and Stock Concentrations for some of the reagents used in Series 3

Reagent Composition
Miniprep Solution 1* 50 mM glucose
10 mM EDTA (Ethylenediaminetetraacetic acid)
25 mM Tris, pH 8
Miniprep Solution 2* 0.2 M NaOH
1% SDS (sodium dodecyl sulfate)
Miniprep Solution 3* 2.7M Potassium Acetate
6.6M Acetic Acid
Luria-Bertoni Medium (LB) 1% Tryptone
0.5% Yeast Extract
1% NaCl
(2% agar in plates)
Nematode Growth Medium Lite(NGM lite)- Available in powder form from US Biological Cat. #N1005 (http://www.usbio.net) 0.3% NaCl
1.7% Bacto-agar
0.25% Bacto-peptone
0.1% (vol/vol)cholesterol
Chemotaxis media 2% bactoagar
5 mM potassium phosphate buffer, pH = 6.0
1 mM CaCl2
1 mM MgSO4
Ampicillin* Stock = 50 mg/ml in water
use 1:1000 in liquid cultures
1:500 in agar
IPTG Stock = 0.5 M
use 0.5 mM in cultures
1% agarose gel 1% (w/v) agarose in 1X TAE buffer. (50X TAE Stock Solution= 242 g/L Tris Base (MW=121.1); 57.1 mL/L Glacial Acetic Acid; 100 mL/L 0.5 M EDTA; Add ddH2O to 1 Liter and adjust pH to 8.5 using KOH)
DNA loading dye 0.4 % Ficoll 400, 1.8 mM EDTA, 0.55 mM Tris-HCl, 0.00117 % SDS, 0.025 % Bromophenol Blue pH 8.0 @ 25°C
1x PBST (phosphate buffered saline with Tween20) 137 mM NaCl,2.7 mM KCl,4.3 mM Na2HPO4,1.47 mM KH2PO4, 0.25% -Adjust to a final pH of 7.4.
50X TAE (use at 1X) 2M Tris Base, 0.05M EDTA, 5.7% Glacial Acetic Acid, adjust to a final pH of 8.5 with KOH
*NOTE: These concentrations are STOCK concentrations - not effective concentrations!
Personal tools