Carrico:Recipes: Difference between revisions
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===2XYT Media (to make 1L)=== | <font face="garamond" size="4" style="color:#000000"> | ||
==='''2XYT Media (to make 1L)'''=== | |||
#Combine: 900mL distilled water, 16g Bacto Tryptone, 10g Bacto Yeast Extract, 5g NaCl | #Combine: 900mL distilled water, 16g Bacto Tryptone, 10g Bacto Yeast Extract, 5g NaCl | ||
| Line 8: | Line 10: | ||
#Autoclave | #Autoclave | ||
==DNA Gel Electrophoresis== | =='''DNA Gel Electrophoresis'''== | ||
===50x TAE Buffer (to make 1L)=== | ==='''50x TAE Buffer (to make 1L)'''=== | ||
*242g 2M Tris base | *242g 2M Tris base | ||
| Line 17: | Line 19: | ||
*Add water up to 1L | *Add water up to 1L | ||
Note: To dilute 50x to 1x, add 20mL of 50x to 980mL water | '''Note:''' To dilute 50x to 1x, add 20mL of 50x to 980mL water | ||
=='''Protein'''== | |||
==='''10x Laemmli Running Buffer (Tris glycine)'''=== | |||
*30.3g Tris base | |||
*144g glycine | |||
*10g SDS | |||
*Add water to 1L | |||
==='''Coomassie Brilliant Blue (CBB)'''=== | |||
#Dissolve 0.25g CBB in 90mL methanol:water (1:1 v/v) and 10mL glacial acetic acid | |||
#Filter through Whatman #1 filter paper to remove particulate matter | |||
#Stain gel for at least 4 hours in 5 volumes of CBB | |||
==='''Protein Gel Destain Solution'''=== | |||
*125mL methanol | |||
*35mL glacial acetic acid | |||
*Add water to 500mL | |||
*Store at room temperature | |||
*Destain for up to 24 hours | |||
Latest revision as of 09:02, 17 July 2009
Carrico Lab at Stony Brook
Home Lab Members Recipes Standard Protocols Publications Links
2XYT Media (to make 1L)
- Combine: 900mL distilled water, 16g Bacto Tryptone, 10g Bacto Yeast Extract, 5g NaCl
- pH to 7.0
- Add water up to 1L
- Autoclave
DNA Gel Electrophoresis
50x TAE Buffer (to make 1L)
- 242g 2M Tris base
- 57.1mL 1M glacial acetic acid
- 200mL 100mM EDTA
- Add water up to 1L
Note: To dilute 50x to 1x, add 20mL of 50x to 980mL water
Protein
10x Laemmli Running Buffer (Tris glycine)
- 30.3g Tris base
- 144g glycine
- 10g SDS
- Add water to 1L
Coomassie Brilliant Blue (CBB)
- Dissolve 0.25g CBB in 90mL methanol:water (1:1 v/v) and 10mL glacial acetic acid
- Filter through Whatman #1 filter paper to remove particulate matter
- Stain gel for at least 4 hours in 5 volumes of CBB
Protein Gel Destain Solution
- 125mL methanol
- 35mL glacial acetic acid
- Add water to 500mL
- Store at room temperature
- Destain for up to 24 hours
