Carrico:Recipes

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===2XYT Media (to make 1L)===
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==='''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
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#Autoclave
#Autoclave
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==DNA Gel Electrophoresis==
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=='''DNA Gel Electrophoresis'''==
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===50x TAE Buffer (to make 1L)===
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==='''50x TAE Buffer (to make 1L)'''===
*242g 2M Tris base
*242g 2M Tris base
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*Add water up to 1L
*Add water up to 1L
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Note: To dilute 50x to 1x, add 20mL of 50x to 980mL water
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'''Note:''' To dilute 50x to 1x, add 20mL of 50x to 980mL water
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=='''Protein'''==
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==='''10x Laemmli Running Buffer (Tris glycine)'''===
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*30.3g Tris base
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*144g glycine
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*10g SDS
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*Add water to 1L
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==='''Coomassie Brilliant Blue (CBB)'''===
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#Dissolve 0.25g CBB in 90mL methanol:water (1:1 v/v) and 10mL glacial acetic acid
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#Filter through Whatman #1 filter paper to remove particulate matter
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#Stain gel for at least 4 hours in 5 volumes of CBB
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==='''Protein Gel Destain Solution'''===
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*125mL methanol
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*35mL glacial acetic acid
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*Add water to 500mL
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*Store at room temperature
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*Destain for up to 24 hours

Current revision


Carrico Lab at Stony Brook


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Contents

2XYT Media (to make 1L)

  1. Combine: 900mL distilled water, 16g Bacto Tryptone, 10g Bacto Yeast Extract, 5g NaCl
  2. pH to 7.0
  3. Add water up to 1L
  4. 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)

  1. Dissolve 0.25g CBB in 90mL methanol:water (1:1 v/v) and 10mL glacial acetic acid
  2. Filter through Whatman #1 filter paper to remove particulate matter
  3. 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
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