Griffitts:Stock solutions

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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

@@ Line 1: / Line 1: @@
-{{Griffitts}}
+<div class="noprint">
+{{Template:Griffitts}}
+</div>
 ==Antibiotics==
 ===Ampicillin (40 mL)===
+For a stock solution of 25 mg/mL:
 * 1 g [[Wikipedia:Ampicillin|Ampicillin]]
 * 40 mL ddH<sub>2</sub>O
 * Filter sterilize
-** Store in refrigerator
+NOTE: Store at 4°C
 ===Chloramphenicol (40 mL)===
-* 0.3 g [[Wikipedia:Chloramphenicol|Chloramphenicol]]
+For a stock solution of 7.5 mg/mL:
-* 40 mL [[Wikipedia:Ethanol|Ethanol]]
+* 300 mg [[Wikipedia:Chloramphenicol|Chloramphenicol]]
-** Do '''not''' filter sterilize
+* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
-** Store in refrigerator
+NOTE: Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter<br>
-===Gentamycin (40 mL)===
+NOTE: Store at 4°C
-* 0.3 g [[Wikipedia:Gentamycin|Gentamycin]]
+===Erythromycin (20 mL)===
+For a stock solution of 10 mg/mL:
+* 200 mg [[Wikipedia:Erythromycin|Erythromycin]]
+* 20 mL 100% ethanol (in fume hood)
+* Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter
+NOTE: Store at &ndash;20°C
+===Gentamicin (40 mL)===
+For a stock solution of 7.5 mg/mL:
+* 300 mg [[Wikipedia:Gentamicin|Gentamicin]]
 * 40 mL ddH<sub>2</sub>O
 * Filter sterilize
-** Store in refrigerator
+NOTE: Store at 4°C
 ===Hygromycin (40 mL)===
+For a stock solution of 5 mg/mL:
+* 200 mg [[Wikipedia:Hygromycin|Hygromycin]]
+* 40 mL ddH<sub>2</sub>O
+* Filter sterilize
+NOTE: Store at 4°C
 ===Kanamycin (40 mL)===
-* 0.1 g [[Wikipedia:Kanamycin|Kanamycin]]
+For a stock solution of 7.5 mg/mL:
+* 300 mg [[Wikipedia:Kanamycin|Kanamycin]]
 * 40 mL ddH<sub>2</sub>O
 * Filter sterilize
-** Store in refrigerator
+NOTE: Store at 4°C
+===Nalidixic acid (30 mL)===
+For a stock solution of 30 mg/mL:
+* 900 mg [[Wikipedia:Nalidixic Acid|Nalidixic Acid]]
+* 360 mg NaOH
+* 30 mL ddH<sub>2</sub>O
+* Filter sterilize
+NOTE: Store at 4°C
 ===Neomycin (40 mL)===
-* 0.8 g [[Wikipedia:Neomycin|Neomycin]]
+For a stock solution of 25 mg/mL:
+* 1 g [[Wikipedia:Neomycin|Neomycin]]
 * 40 mL ddH<sub>2</sub>O
 * Filter sterilize
-** Store in refrigerator
+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
+* Filter sterilize
+NOTE: Store at 4°C
+===Rifampicin (20 mL)===
+For a stock solution of 50 mg/mL:
+* 1 g [[Wikipedia:Rifampicin|Rifampicin]]
+* 20 mL N,N-dimethylformamide (DMF; in fume hood)
+NOTE: Do '''not''' filter sterilize&mdash;the DMF will dissolve the filter<br>
+NOTE: Store at &ndash;20°C
 ===Spectinomycin (40 mL)===
-* 20 mg [[Wikipedia:Spectinomycin|Spectinomycin]]
+For a stock solution of 20 mg/mL:
+* 800 mg [[Wikipedia:Spectinomycin|Spectinomycin]]
 * 40 mL ddH<sub>2</sub>O
 * Filter sterilize
-** Store in refrigerator
+NOTE: Store at 4°C
 ===Streptomycin (40 mL)===
-* 2.0 g [[Wikipedia:Streptomycin|Streptomycin]]
+For a stock solution of 50 mg/mL:
+* 2 g [[Wikipedia:Streptomycin|Streptomycin]]
 * 40 mL ddH<sub>2</sub>O
 * Filter sterilize
-** Store in refrigerator
+NOTE: Store at 4°C
 ===Tetracycline (40 mL)===
-* 0.1 g [[Wikipedia:Tetracycline|Tetracycline]]
+For a stock solution of 2.5 mg/mL:
-* 40 mL [[Wikipedia:Ethanol|Ethanol]]
+* 100 mg [[Wikipedia:Tetracycline|Tetracycline]]
-** Do '''not''' filter sterilize
+* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
-** Store in refrigerator
+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
+==Antibiotics Table==
+{| border="1" cellpadding="5" cellspacing="0" class="wikitable sortable"
+|+'''Antibiotic Specifications'''
+|-
+! width="120" style="background:#efefef;" | Antibiotic
+! width="30" style="background:#efefef;" | Abbr.
+! width="90" style="background:#efefef;" | Stripe
+! width="90" style="background:#efefef;" | Stock
+! width="120" style="background:#efefef;" | ''E. coli''
+! width="120" style="background:#efefef;" | ''S. meliloti''
+! width="90" style="background:#efefef;" | Solvent
+! width="90" style="background:#efefef;" | Storage
+|-
+| Ampicillin
+| Ap
+! style="background:#000000;" | <font color=white>'''BLACK'''</font>
+| 25 mg/mL
+| 100 μg/mL (250X)
+| N/A
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Chloramphenicol
+| Cm
+! style="background:#8B4513;" | <font color=white>'''BROWN'''</font>
+| 7.5 mg/mL
+| 30 μg/mL (250X)
+| N/A
+| 50% EtOH
+| 4°C
+|-
+| Erythromycin
+| Er
+| <center>(none)</center>
+| 10 mg/mL
+| N/A
+| N/A
+| 100% EtOH
+| &ndash;20°C
+|-
+| Gentamycin
+| Gm
+! style="background:#9400D3;" | <font color=white>'''PURPLE'''</font>
+| 7.5 mg/mL
+| 3 μg/mL (2500X)
+| 15 μg/mL (500X)
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Hygromycin
+| Hy
+| <center>(none)</center>
+| 5 mg/mL
+| 20 μg/mL (250X)
+| 20 μg/mL (250X)
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Kanamycin
+| Km
+! style="background:#FFD700;" | <font color=white>'''YELLOW'''</font>
+| 7.5 mg/mL
+| 30 μg/mL (250X)
+| '''Use Neomycin'''
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Nalidixic acid
+| Na
+| <center>(none)</center>
+| 30 mg/mL
+| 30 μg/mL (1000X)
+| N/A
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Neomycin
+| Nm
+! 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
+| N/A
+| 100 μg/mL (250X)
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Rifampicin
+| Rf
+| <center>(none)</center>
+| 50 mg/mL
+| N/A
+| 100 μg/mL (500X)
+| DMF
+| &ndash;20°C
+|-
+| Spectinomycin
+| Sp
+! style="background:#FF8C00;" | <font color=white>'''ORANGE'''</font>
+| 20 mg/mL
+| 80 μg/mL (250X)
+| 80 μg/mL (250X)
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Streptomycin
+| Sm
+! style="background:#0000FF;" | <font color=white>'''BLUE'''</font>
+| 50 mg/mL
+| N/A
+| 200 μg/mL (250X)
+| ddH<sub>2</sub>O
+| 4°C
+|-
+| Tetracycline
+| Tc
+! style="background:#FF0000;" | <font color=white>'''RED'''</font>
+| 2.5 mg/mL
+| 5 μg/mL (500X)
+| 5 μg/mL (500X)
+| 50% EtOH
+| 4°C
+|-
+| Triclosan
+| Tr
+| <center>(none)</center>
+| 10 mg/mL
+| N/A
+| N/A
+| 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
+|}
+Note: These parameters are intended to be compatible with pre-made 250-mL amounts of [[Griffitts:Common media#LB Broth (1 L)|LB]] agar, such that usually 1 mL of stock solution is diluted into the [[Griffitts:Common media#LB Broth (1 L)|LB]] agar. For liquid [[Griffitts:Common media#LB Broth (1 L)|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 100 mM stock solutions of G, A, T, C (NEB)
+* Thaw and mix (stir) 100 mM stock solutions of dGTP, dATP, dTTP, dCTP (NEB; in freezer)
-* Combine in a 1.5-mL tube:
+* Master Mix:
-** 60 μl H<sub>2</sub>O
+** 60 μl sterile dH<sub>2</sub>O
-** 10 μl [[Wikipedia:Guanine|G]]
+** 10 μl [[Wikipedia:Guanine|dGTP]]
-** 10 μl [[Wikipedia:Adenine|A]]
+** 10 μl [[Wikipedia:Adenine|dATP]]
-** 10 μl [[Wikipedia:Thymine|T]]
+** 10 μl [[Wikipedia:Thymine|dTTP]]
-** 10 μl [[Wikipedia:Cytosine|C]]
+** 10 μl [[Wikipedia:Cytosine|dCTP]]
+* Dispense 100-μL aliquots into 1.7-mL tubes
+Note: Store in the &ndash;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&mdash;do '''not''' dilute!
+Note: Store in the &ndash;20°C freezer in the box marked "Other Enzymes"
 ===10X ''Pfx'' buffer===
-===20X Triton X-100===
+* 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&mdash;do '''not''' dilute!
+Note: Store in the &ndash;20°C freezer in the box marked "Other Enzymes"
+===8X DNA loading===
+* 40% [[Griffitts:Stock solutions#80% Glycerol (75 mL)|glycerol]]
+* 1/50 dilution of [[Griffitts:Common buffers#2% bromophenol blue|2% bromophenol blue]]
+===DNA ladder===
+* 140 μL ddH<sub>2</sub>0
+* 40 μL [[Griffitts:Common buffers#8X DNA loading|8X DNA loading]] buffer
+* 20 μL 1 kb ladder from NEB
+* 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
+===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)===
+* 10 mL ddH<sub>2</sub>O
+* 10 mg ethidium bromide<br>
+Note: use 8 μL per 50 mL gel<br>
+WARNING: Always wear gloves when handling ethidium bromide
 ==Media Additives==
 ===1 M Calcium Chloride (75 mL)===
-* 75 mL dH<sub>2</sub>O
+* 75 mL ddH<sub>2</sub>O
-* 11.027 g CaCl<sub>2</sub>·2H<sub>2</sub>O
+* 11.027 g calcium chloride dihydrate (CaCl<sub>2</sub>·2H<sub>2</sub>O)
 Autoclave
-===X-gal===
+===2 M Magnesium Chloride (75 mL)===
-===IPTG===
+* 75 mL ddH<sub>2</sub>O
-===Copper sulfate===
+* 30.4965 g magnesium chloride hexahydrate (MgCl<sub>2</sub>·6H<sub>2</sub>O)
-===SDS===
+Autoclave
+===1 M Sodium Citrate (40 mL)===
+* 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)
+* pH to 5.7 with HCl (~1 mL)
+* QS to 40 mL with ddH<sub>2</sub>O
+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 &ndash;20°C
+===0.2 M IPTG===
+* 5 mL ddH<sub>2</sub>O
+* 0.238g isopropyl-β-D-thiogalactopyranoside (IPTG)(in freezer)
+Sterile filter, aliquot, and store at &ndash;20°C
+===1 M Copper (II) sulfate (50 mL)===
+* 50 mL ddH<sub>2</sub>O
+* 7.98 g CuSO<sub>4</sub>
+Sterile filter
+===10% [[SDS]] (75 mL)===
+* 75 mL ddH<sub>2</sub>O
+* 7.5 g Sodium dodecyl sulfate ([[SDS]])
+Do '''not''' sterilize
+===20% [[SDS]] (75 mL)===
+* 75 mL ddH<sub>2</sub>O
+* 15 g Sodium dodecyl sulfate ([[SDS]])
+Do '''not''' sterilize
+===30% Acrylamide (75 mL)===
+* 75 mL ddH<sub>2</sub>O
+* 22.5 g acrylamide
+===40% Acrylamide (75 mL)===
+* 75 mL ddH<sub>2</sub>O
+* 30 g acrylamide
+===10% Ammonium persulfate (75 mL)===
+* 75 mL ddH<sub>2</sub>O
+* 7.5 g ammonium persulfate (APS)
+===5 M NaCl (75 mL)===
+* 75 mL ddH<sub>2</sub>O
+* 21.92 g NaCl
+Autoclave
+===80% Glycerol (75 mL)===
+* 15 mL ddH<sub>2</sub>O
+* 60 mL glycerol
+Microwave for 30-second intervals on high until dissolved<br>
+Autoclave
+===1 M DTT (5 mL) ===
+* 5 mL ddH<sub>2</sub>O
+* 771 mg Dithiothreitol (DTT; in fridge)
+Aliquot 500 μL into Eppendorf tubes
+Store in &ndash;20°C
+===Orcinol spray===
+* 5 mL ddH<sub>2</sub>O
+* 25 mL methanol (in fume hood)
+* 10 mL sulfuric acid (H<sub>2</sub>SO<sub>4</sub> (in fume hood&mdash;be careful!)
+* 200 mg orcinol

Griffitts:Stock solutions

From OpenWetWare

Revision as of 18:41, 29 February 2012

Contents

Antibiotics

Ampicillin (40 mL)

Chloramphenicol (40 mL)

Erythromycin (20 mL)

Gentamicin (40 mL)

Hygromycin (40 mL)

Kanamycin (40 mL)

Nalidixic acid (30 mL)

Neomycin (40 mL)

Phosphomycin

Rifampicin (20 mL)

Spectinomycin (40 mL)

Streptomycin (40 mL)

Tetracycline (40 mL)

Triclosan (40 mL)

Trimethoprim (40 mL)

Antibiotics Table

DNA

10 mM dNTPs

10X Taq buffer

10X Pfx buffer

8X DNA loading

DNA ladder

Ethidium Bromide (1 mL)

Ethidium Bromide (10 mL)

Media Additives

1 M Calcium Chloride (75 mL)

2 M Magnesium Chloride (75 mL)

1 M Sodium Citrate (40 mL)

2% X-gal

0.2 M IPTG

1 M Copper (II) sulfate (50 mL)

10% SDS (75 mL)

20% SDS (75 mL)

30% Acrylamide (75 mL)

40% Acrylamide (75 mL)

10% Ammonium persulfate (75 mL)

5 M NaCl (75 mL)

80% Glycerol (75 mL)

1 M DTT (5 mL)

Orcinol spray

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