Griffitts:Stock solutions

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Revision as of 12:40, 17 March 2010 (view source) Matthew B. Crook (Talk | contribs) m (→Antibiotics) ← Previous diff		Revision as of 18:41, 29 February 2012 (view source) Matthew B. Crook (Talk | contribs) m (→Triclosan (40 mL)) Next diff →
(11 intermediate revisions not shown.)
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	==Antibiotics==		==Antibiotics==
	===Ampicillin (40 mL)===		===Ampicillin (40 mL)===
		+	For a stock solution of 25 mg/mL:
	* 1 g [[Wikipedia:Ampicillin|Ampicillin]]		* 1 g [[Wikipedia:Ampicillin|Ampicillin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
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	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Chloramphenicol (40 mL)===		===Chloramphenicol (40 mL)===
		+	For a stock solution of 7.5 mg/mL:
	* 300 mg [[Wikipedia:Chloramphenicol|Chloramphenicol]]		* 300 mg [[Wikipedia:Chloramphenicol|Chloramphenicol]]
	* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]		* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
Line 14:		Line 16:
	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Erythromycin (20 mL)===		===Erythromycin (20 mL)===
-	* 1 g [[Wikipedia:Erythromycin|Erythromycin]]	+	For a stock solution of 10 mg/mL:
		+	* 200 mg [[Wikipedia:Erythromycin|Erythromycin]]
	* 20 mL 100% ethanol (in fume hood)		* 20 mL 100% ethanol (in fume hood)
	* Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter		* Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter
	NOTE: Store at &ndash;20°C		NOTE: Store at &ndash;20°C
	===Gentamicin (40 mL)===		===Gentamicin (40 mL)===
		+	For a stock solution of 7.5 mg/mL:
	* 300 mg [[Wikipedia:Gentamicin|Gentamicin]]		* 300 mg [[Wikipedia:Gentamicin|Gentamicin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
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	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Hygromycin (40 mL)===		===Hygromycin (40 mL)===
		+	For a stock solution of 5 mg/mL:
	* 200 mg [[Wikipedia:Hygromycin|Hygromycin]]		* 200 mg [[Wikipedia:Hygromycin|Hygromycin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
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	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Kanamycin (40 mL)===		===Kanamycin (40 mL)===
		+	For a stock solution of 7.5 mg/mL:
	* 300 mg [[Wikipedia:Kanamycin|Kanamycin]]		* 300 mg [[Wikipedia:Kanamycin|Kanamycin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
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	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Nalidixic acid (30 mL)===		===Nalidixic acid (30 mL)===
		+	For a stock solution of 30 mg/mL:
	* 900 mg [[Wikipedia:Nalidixic Acid|Nalidixic Acid]]		* 900 mg [[Wikipedia:Nalidixic Acid|Nalidixic Acid]]
	* 360 mg NaOH		* 360 mg NaOH
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	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Neomycin (40 mL)===		===Neomycin (40 mL)===
		+	For a stock solution of 25 mg/mL:
	* 1 g [[Wikipedia:Neomycin|Neomycin]]		* 1 g [[Wikipedia:Neomycin|Neomycin]]
		+	* 40 mL ddH<sub>2</sub>O
		+	* Filter sterilize
		+	NOTE: Store at 4°C
		+	===Phosphomycin===
		+	For a stock solution of 25 mg/mL:
		+	* 1 g [[Wikipedia:Fosfomycin|Phosphomycin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
	* Filter sterilize		* Filter sterilize
	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Rifampicin (20 mL)===		===Rifampicin (20 mL)===
		+	For a stock solution of 50 mg/mL:
	* 1 g [[Wikipedia:Rifampicin|Rifampicin]]		* 1 g [[Wikipedia:Rifampicin|Rifampicin]]
	* 20 mL N,N-dimethylformamide (DMF; in fume hood)		* 20 mL N,N-dimethylformamide (DMF; in fume hood)
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	NOTE: Store at &ndash;20°C		NOTE: Store at &ndash;20°C
	===Spectinomycin (40 mL)===		===Spectinomycin (40 mL)===
		+	For a stock solution of 20 mg/mL:
	* 800 mg [[Wikipedia:Spectinomycin|Spectinomycin]]		* 800 mg [[Wikipedia:Spectinomycin|Spectinomycin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
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	NOTE: Store at 4°C		NOTE: Store at 4°C
	===Streptomycin (40 mL)===		===Streptomycin (40 mL)===
		+	For a stock solution of 50 mg/mL:
	* 2 g [[Wikipedia:Streptomycin|Streptomycin]]		* 2 g [[Wikipedia:Streptomycin|Streptomycin]]
	* 40 mL ddH<sub>2</sub>O		* 40 mL ddH<sub>2</sub>O
	* Filter sterilize		* Filter sterilize
	NOTE: Store at 4°C		NOTE: Store at 4°C
		+
	===Tetracycline (40 mL)===		===Tetracycline (40 mL)===
		+	For a stock solution of 2.5 mg/mL:
	* 100 mg [[Wikipedia:Tetracycline|Tetracycline]]		* 100 mg [[Wikipedia:Tetracycline|Tetracycline]]
	* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]		* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
	NOTE: Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter<br>		NOTE: Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter<br>
		+	NOTE: Store at 4°C
		+	===Triclosan (40 mL)===
		+	For a stock solution of 10 mg/mL:
		+	* 400 mg [[Wikipedia:Triclosan|Triclosan]]
		+	* 40 mL 100% [[Wikipedia:Ethanol|Ethanol]]
		+	NOTE: Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter<br>
		+	NOTE: Store at 4°C
		+
		+	===Trimethoprim (40 mL)===
		+	For a stock solution of 6.25 mg/mL:
		+	* 250 mg [[Wikipedia:Trimethoprim|Trimethoprim]]
		+	* 40 mL 100% [[Wikipedia:Dimethyl sulfoxide|DMSO]]
		+	NOTE: Do '''not''' filter sterilize&mdash;the DMSO may dissolve the filter<br>
	NOTE: Store at 4°C		NOTE: Store at 4°C
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	! width="120" style="background:#efefef;" | ''E. coli''		! width="120" style="background:#efefef;" | ''E. coli''
	! width="120" style="background:#efefef;" | ''S. meliloti''		! width="120" style="background:#efefef;" | ''S. meliloti''
-	! width="60" style="background:#efefef;" | Solvent	+	! width="90" style="background:#efefef;" | Solvent
-	! width="60" style="background:#efefef;" | Storage	+	! width="90" style="background:#efefef;" | Storage
	|-		|-
	| Ampicillin		| Ampicillin
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	| 30 μg/mL (250X)		| 30 μg/mL (250X)
	| N/A		| N/A
-	| EtOH	+	| 50% EtOH
	| 4°C		| 4°C
	|-		|-
	| Erythromycin		| Erythromycin
-	| Em	+	| Er
	| <center>(none)</center>		| <center>(none)</center>
-	| 50 mg/mL	+	| 10 mg/mL
	| N/A		| N/A
	| N/A		| N/A
-	| EtOH	+	| 100% EtOH
	| &ndash;20°C		| &ndash;20°C
	|-		|-
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	| 7.5 mg/mL		| 7.5 mg/mL
	| 30 μg/mL (250X)		| 30 μg/mL (250X)
-	| N/A	+	| '''Use Neomycin'''
	| ddH<sub>2</sub>O		| ddH<sub>2</sub>O
	| 4°C		| 4°C
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	| Na		| Na
	| <center>(none)</center>		| <center>(none)</center>
-	| 5 mg/mL	+	| 30 mg/mL
	| 30 μg/mL (1000X)		| 30 μg/mL (1000X)
	| N/A		| N/A
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	| Nm		| Nm
	! style="background:#008000;" | <font color=white>'''GREEN'''</font>		! style="background:#008000;" | <font color=white>'''GREEN'''</font>
		+	| 25 mg/mL
		+	| '''Use Kanamycin'''
		+	| 100 μg/mL (250X)
		+	| ddH<sub>2</sub>O
		+	| 4°C
		+	|-
		+	| Phosphomycin
		+	| Pm
		+	| <center>(none)</center>
	| 25 mg/mL		| 25 mg/mL
	| N/A		| N/A
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	|-		|-
	| Rifampicin		| Rifampicin
-	| Rm	+	| Rf
	| <center>(none)</center>		| <center>(none)</center>
	| 50 mg/mL		| 50 mg/mL
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	! style="background:#FF0000;" | <font color=white>'''RED'''</font>		! style="background:#FF0000;" | <font color=white>'''RED'''</font>
	| 2.5 mg/mL		| 2.5 mg/mL
-	| 10 μg/mL (250X)	+	| 5 μg/mL (500X)
-	| 10 μg/mL (250X)	+	| 5 μg/mL (500X)
-	| EtOH	+	| 50% EtOH
	| 4°C		| 4°C
	|-		|-
	| Triclosan		| Triclosan
-	|	+	| Tr
	| <center>(none)</center>		| <center>(none)</center>
-	|	+	| 10 mg/mL
	| N/A		| N/A
	| N/A		| N/A
-	| EtOH	+	| 100% EtOH
		+	| 4°C
		+	|-
		+	| Trimethoprim
		+	| Tm
		+	| <center>(none)</center>
		+	| 6.25 mg/mL
		+	| N/A
		+	| 12.5 μg/mL (500X)<br><center>or</center>25 μg/mL (250X)
		+	| 100% DMSO
	| 4°C		| 4°C
	|}		|}
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	* 1/50 dilution of [[Griffitts:Common buffers#2% bromophenol blue|2% bromophenol blue]]		* 1/50 dilution of [[Griffitts:Common buffers#2% bromophenol blue|2% bromophenol blue]]
	===DNA ladder===		===DNA ladder===
-	* 147 μL ddH<sub>2</sub>0	+	* 140 μL ddH<sub>2</sub>0
-	* 34 μL [[Griffitts:Common buffers#8X DNA loading|8X DNA loading]] buffer	+	* 40 μL [[Griffitts:Common buffers#8X DNA loading|8X DNA loading]] buffer
	* 20 μL 1 kb ladder from NEB		* 20 μL 1 kb ladder from NEB
	* 2 μL [[Griffitts:Common buffers#0.25 M EDTA (75 mL)|0.25 M EDTA]]<br>		* 2 μL [[Griffitts:Common buffers#0.25 M EDTA (75 mL)|0.25 M EDTA]]<br>
	Note: use 5 μL per lane; 3kb band = 60 ng		Note: use 5 μL per lane; 3kb band = 60 ng
		+
		+	===Ethidium Bromide (1 mL)===
		+	* 100 μL 10 mg/mL ethidium bromide
		+	** This is stored on the middle shelf of the refrigerator on the right
		+	* 900 μL ddH<sub>2</sub>O
		+	Note: use 8 μL per 50 mL gel<br>
		+	WARNING: Always wear gloves when handling ethidium bromide
		+
	===Ethidium Bromide (10 mL)===		===Ethidium Bromide (10 mL)===
-	* 10 mL ddH<sub>2</sub>0	+	* 10 mL ddH<sub>2</sub>O
	* 10 mg ethidium bromide<br>		* 10 mg ethidium bromide<br>
-	Note: use 8 μL per 50 mL gel	+	Note: use 8 μL per 50 mL gel<br>
		+	WARNING: Always wear gloves when handling ethidium bromide
	==Media Additives==		==Media Additives==
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	===1 M Sodium Citrate (40 mL)===		===1 M Sodium Citrate (40 mL)===
-	* 40 mL ddH<sub>2</sub>O	+	* 35 mL ddH<sub>2</sub>O
	* 11.76  g sodium citrate tribasic dihydrate (Na<sub>3</sub>C<sub>6</sub>H<sub>5</sub>O<sub>7</sub>·2H<sub>2</sub>O)		* 11.76  g sodium citrate tribasic dihydrate (Na<sub>3</sub>C<sub>6</sub>H<sub>5</sub>O<sub>7</sub>·2H<sub>2</sub>O)
-	* pH to 5.7 with HCl	+	* pH to 5.7 with HCl (~1 mL)
		+	* QS to 40 mL with ddH<sub>2</sub>O
	Sterile filter		Sterile filter

---

Revision as of 18:41, 29 February 2012

Contents 1 Antibiotics 1.1 Ampicillin (40 mL) 1.2 Chloramphenicol (40 mL) 1.3 Erythromycin (20 mL) 1.4 Gentamicin (40 mL) 1.5 Hygromycin (40 mL) 1.6 Kanamycin (40 mL) 1.7 Nalidixic acid (30 mL) 1.8 Neomycin (40 mL) 1.9 Phosphomycin 1.10 Rifampicin (20 mL) 1.11 Spectinomycin (40 mL) 1.12 Streptomycin (40 mL) 1.13 Tetracycline (40 mL) 1.14 Triclosan (40 mL) 1.15 Trimethoprim (40 mL) 2 Antibiotics Table 3 DNA 3.1 10 mM dNTPs 3.2 10X Taq buffer 3.3 10X Pfx buffer 3.4 8X DNA loading 3.5 DNA ladder 3.6 Ethidium Bromide (1 mL) 3.7 Ethidium Bromide (10 mL) 4 Media Additives 4.1 1 M Calcium Chloride (75 mL) 4.2 2 M Magnesium Chloride (75 mL) 4.3 1 M Sodium Citrate (40 mL) 4.4 2% X-gal 4.5 0.2 M IPTG 4.6 1 M Copper (II) sulfate (50 mL) 4.7 10% SDS (75 mL) 4.8 20% SDS (75 mL) 4.9 30% Acrylamide (75 mL) 4.10 40% Acrylamide (75 mL) 4.11 10% Ammonium persulfate (75 mL) 4.12 5 M NaCl (75 mL) 4.13 80% Glycerol (75 mL) 4.14 1 M DTT (5 mL) 4.15 Orcinol spray

Antibiotics

Ampicillin (40 mL)

For a stock solution of 25 mg/mL:

• 1 g Ampicillin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Chloramphenicol (40 mL)

For a stock solution of 7.5 mg/mL:

• 300 mg Chloramphenicol
• 40 mL 50% Ethanol

NOTE: Do not filter sterilize—the ethanol may dissolve the filter
NOTE: Store at 4°C

Erythromycin (20 mL)

For a stock solution of 10 mg/mL:

• 200 mg Erythromycin
• 20 mL 100% ethanol (in fume hood)
• Do not filter sterilize—the ethanol may dissolve the filter

NOTE: Store at –20°C

Gentamicin (40 mL)

For a stock solution of 7.5 mg/mL:

• 300 mg Gentamicin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Hygromycin (40 mL)

For a stock solution of 5 mg/mL:

• 200 mg Hygromycin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Kanamycin (40 mL)

For a stock solution of 7.5 mg/mL:

• 300 mg Kanamycin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Nalidixic acid (30 mL)

For a stock solution of 30 mg/mL:

• 900 mg Nalidixic Acid
• 360 mg NaOH
• 30 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Neomycin (40 mL)

For a stock solution of 25 mg/mL:

• 1 g Neomycin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Phosphomycin

For a stock solution of 25 mg/mL:

• 1 g Phosphomycin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Rifampicin (20 mL)

For a stock solution of 50 mg/mL:

• 1 g Rifampicin
• 20 mL N,N-dimethylformamide (DMF; in fume hood)

NOTE: Do not filter sterilize—the DMF will dissolve the filter
NOTE: Store at –20°C

Spectinomycin (40 mL)

For a stock solution of 20 mg/mL:

• 800 mg Spectinomycin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Streptomycin (40 mL)

For a stock solution of 50 mg/mL:

• 2 g Streptomycin
• 40 mL ddH₂O
• Filter sterilize

NOTE: Store at 4°C

Tetracycline (40 mL)

For a stock solution of 2.5 mg/mL:

• 100 mg Tetracycline
• 40 mL 50% Ethanol

NOTE: Do not filter sterilize—the ethanol may dissolve the filter
NOTE: Store at 4°C

Triclosan (40 mL)

For a stock solution of 10 mg/mL:

• 400 mg Triclosan
• 40 mL 100% Ethanol

NOTE: Do not filter sterilize—the ethanol may dissolve the filter
NOTE: Store at 4°C

Trimethoprim (40 mL)

For a stock solution of 6.25 mg/mL:

• 250 mg Trimethoprim
• 40 mL 100% DMSO

NOTE: Do not filter sterilize—the DMSO may dissolve the filter
NOTE: Store at 4°C

Antibiotics Table

Antibiotic Specifications

Antibiotic	Abbr.	Stripe	Stock	E. coli	S. meliloti	Solvent	Storage
Ampicillin	Ap	BLACK	25 mg/mL	100 μg/mL (250X)	N/A	ddH2O	4°C
Chloramphenicol	Cm	BROWN	7.5 mg/mL	30 μg/mL (250X)	N/A	50% EtOH	4°C
Erythromycin	Er	(none)	10 mg/mL	N/A	N/A	100% EtOH	–20°C
Gentamycin	Gm	PURPLE	7.5 mg/mL	3 μg/mL (2500X)	15 μg/mL (500X)	ddH2O	4°C
Hygromycin	Hy	(none)	5 mg/mL	20 μg/mL (250X)	20 μg/mL (250X)	ddH2O	4°C
Kanamycin	Km	YELLOW	7.5 mg/mL	30 μg/mL (250X)	Use Neomycin	ddH2O	4°C
Nalidixic acid	Na	(none)	30 mg/mL	30 μg/mL (1000X)	N/A	ddH2O	4°C
Neomycin	Nm	GREEN	25 mg/mL	Use Kanamycin	100 μg/mL (250X)	ddH2O	4°C
Phosphomycin	Pm	(none)	25 mg/mL	N/A	100 μg/mL (250X)	ddH2O	4°C
Rifampicin	Rf	(none)	50 mg/mL	N/A	100 μg/mL (500X)	DMF	–20°C
Spectinomycin	Sp	ORANGE	20 mg/mL	80 μg/mL (250X)	80 μg/mL (250X)	ddH2O	4°C
Streptomycin	Sm	BLUE	50 mg/mL	N/A	200 μg/mL (250X)	ddH2O	4°C
Tetracycline	Tc	RED	2.5 mg/mL	5 μg/mL (500X)	5 μg/mL (500X)	50% EtOH	4°C
Triclosan	Tr	(none)	10 mg/mL	N/A	N/A	100% EtOH	4°C
Trimethoprim	Tm	(none)	6.25 mg/mL	N/A	12.5 μg/mL (500X) or25 μg/mL (250X)	100% DMSO	4°C

Note: These parameters are intended to be compatible with pre-made 250-mL amounts of LB agar, such that usually 1 mL of stock solution is diluted into the LB agar. For liquid LB (typically 75 mL amounts) 0.3 mL of the stock solution will usually be added. Exceptions: Gentamycin, Nalidixic Acid, and Rifampicin.

DNA

10 mM dNTPs

• Thaw and mix (stir) 100 mM stock solutions of dGTP, dATP, dTTP, dCTP (NEB; in freezer)
• Master Mix:
  • 60 μl sterile dH₂O
  • 10 μl dGTP
  • 10 μl dATP
  • 10 μl dTTP
  • 10 μl dCTP
• Dispense 100-μL aliquots into 1.7-mL tubes

Note: Store in the –20°C freezer in the box marked "Other Enzymes"

10X Taq buffer

• Label tubes: 10X Taq buffer
• Thaw and mix (stir) "Thermo pol buffer" (NEB; in freezer)
• Aliquot 375 μL of "Thermo pol buffer" into tubes—do not dilute!

Note: Store in the –20°C freezer in the box marked "Other Enzymes"

10X Pfx buffer

• Label tubes: 10X Pfx buffer
• Thaw and mix (stir) "Pfx50 10X PCR Buffer" (NEB; in freezer)
• Aliquot 375 μL of "Pfx50 10X PCR Buffer" into tubes—do not dilute!

Note: Store in the –20°C freezer in the box marked "Other Enzymes"

8X DNA loading

• 40% glycerol
• 1/50 dilution of 2% bromophenol blue

DNA ladder

• 140 μL ddH₂0
• 40 μL 8X DNA loading buffer
• 20 μL 1 kb ladder from NEB
• 2 μL 0.25 M EDTA
  

Note: use 5 μL per lane; 3kb band = 60 ng

Ethidium Bromide (1 mL)

• 100 μL 10 mg/mL ethidium bromide
  • This is stored on the middle shelf of the refrigerator on the right
• 900 μL ddH₂O

Note: use 8 μL per 50 mL gel
WARNING: Always wear gloves when handling ethidium bromide

Ethidium Bromide (10 mL)

• 10 mL ddH₂O
• 10 mg ethidium bromide
  

Note: use 8 μL per 50 mL gel
WARNING: Always wear gloves when handling ethidium bromide

Media Additives

1 M Calcium Chloride (75 mL)

• 75 mL ddH₂O
• 11.027 g calcium chloride dihydrate (CaCl₂·2H₂O)

Autoclave

2 M Magnesium Chloride (75 mL)

• 75 mL ddH₂O
• 30.4965 g magnesium chloride hexahydrate (MgCl₂·6H₂O)

Autoclave

1 M Sodium Citrate (40 mL)

• 35 mL ddH₂O
• 11.76 g sodium citrate tribasic dihydrate (Na₃C₆H₅O₇·2H₂O)
• pH to 5.7 with HCl (~1 mL)
• QS to 40 mL with ddH₂O

Sterile filter

2% X-gal

• 10 mL N,N-dimethylformamide (DMF)(in fume hood)
• 0.2 g 5-bromo-4-chloro-3-indolyl- beta-D-galactopyranoside (X-gal)(in freezer)

Store at –20°C

0.2 M IPTG

• 5 mL ddH₂O
• 0.238g isopropyl-β-D-thiogalactopyranoside (IPTG)(in freezer)

Sterile filter, aliquot, and store at –20°C

1 M Copper (II) sulfate (50 mL)

• 50 mL ddH₂O
• 7.98 g CuSO₄

Sterile filter

10% SDS (75 mL)

• 75 mL ddH₂O
• 7.5 g Sodium dodecyl sulfate (SDS)

Do not sterilize

20% SDS (75 mL)

• 75 mL ddH₂O
• 15 g Sodium dodecyl sulfate (SDS)

Do not sterilize

30% Acrylamide (75 mL)

• 75 mL ddH₂O
• 22.5 g acrylamide

40% Acrylamide (75 mL)

• 75 mL ddH₂O
• 30 g acrylamide

10% Ammonium persulfate (75 mL)

• 75 mL ddH₂O
• 7.5 g ammonium persulfate (APS)

5 M NaCl (75 mL)

• 75 mL ddH₂O
• 21.92 g NaCl

Autoclave

80% Glycerol (75 mL)

• 15 mL ddH₂O
• 60 mL glycerol

Microwave for 30-second intervals on high until dissolved
Autoclave

1 M DTT (5 mL)

• 5 mL ddH₂O
• 771 mg Dithiothreitol (DTT; in fridge)

Aliquot 500 μL into Eppendorf tubes Store in –20°C

Orcinol spray

• 5 mL ddH₂O
• 25 mL methanol (in fume hood)
• 10 mL sulfuric acid (H₂SO₄ (in fume hood—be careful!)
• 200 mg orcinol