View source for Griffitts:Stock solutions

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==Antibiotics==
===Ampicillin (40 mL)===
* 1 g [[Wikipedia:Ampicillin|Ampicillin]]
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator
===Chloramphenicol (40 mL)===
* 0.3 g [[Wikipedia:Chloramphenicol|Chloramphenicol]]
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
** Do '''not''' filter sterilize
** Store in refrigerator
===Erythromycin (20 mL)===
* 1.0 g [[Wikipedia:Erythromycin|Erythromycin]]
* 20 mL 100% ethanol (in fume hood)
* Do '''not''' filter sterilize&mdash;the ethanol may dissolve the filter
** Store in the -20°C Freezer

===Gentamycin (40 mL)===
* 0.3 g [[Wikipedia:Gentamycin|Gentamycin]]
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator

===Hygromycin (40 mL)===
===Kanamycin (40 mL)===
* 0.3 g [[Wikipedia:Kanamycin|Kanamycin]]
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator

===Nalidixic acid (30 mL)===
* 0.9 g [[Wikipedia:Nalidixic Acid|Nalidixic Acid]]
* 0.36 g NaOH
* 30 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator

===Neomycin (40 mL)===
* 0.8 g [[Wikipedia:Neomycin|Neomycin]]
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator

===Rifampicin (20 mL)===
* 1.0 g [[Wikipedia:Rifampicin|Rifampicin]]
* 20 mL N,N-dimethylformamide (DMF; in fume hood)
* Do '''not''' filter sterilize&mdash;the DMF will dissolve the filter
** Store in the -20°C Freezer

===Spectinomycin (40 mL)===
* 0.8 g [[Wikipedia:Spectinomycin|Spectinomycin]]
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator

===Streptomycin (40 mL)===
* 2.0 g [[Wikipedia:Streptomycin|Streptomycin]]
* 40 mL ddH<sub>2</sub>O
* Filter sterilize
** Store in refrigerator
===Tetracycline (40 mL)===
* 0.1 g [[Wikipedia:Tetracycline|Tetracycline]]
* 40 mL 50% [[Wikipedia:Ethanol|Ethanol]]
** Do '''not''' filter sterilize
** Store in refrigerator

==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="60" style="background:#efefef;" | Solvent
! width="60" 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
| EtOH
| 4°C
|-
| Erythromycin
| Em
| (none)
| 50 mg/mL
| N/A
| N/A
| 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
| (none)
| 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)
| N/A
| ddH<sub>2</sub>O
| 4°C
|-
| Nalidixic acid
| Na
| (none)
| 5 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>
| 20 mg/mL
| N/A
| 80 μg/mL (250X)
| ddH<sub>2</sub>O
| 4°C
|-
| Rifampicin
| Rm
| (none)
| 50 mg/mL
| N/A
| N/A
| 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
| 10 μg/mL (250X)
| 10 μg/mL (250X)
| EtOH
| 4°C
|-
| Triclosan
| 
| (none)
| 
| N/A
| N/A
| EtOH
| 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. Exception: Gentamycin for ''E. coli''.

==DNA==
===10 mM dNTPs===
* Thaw and mix (stir) 100 mM stock solutions of dGTP, dATP, dTTP, dCTP (NEB; in freezer)
* Combine in a 1.5-mL tube:
** 60 μl H<sub>2</sub>O
** 10 μl [[Wikipedia:Guanine|dGTP]]
** 10 μl [[Wikipedia:Adenine|dATP]]
** 10 μl [[Wikipedia:Thymine|dTTP]]
** 10 μl [[Wikipedia:Cytosine|dCTP]]
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&mdash;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&mdash;do '''not''' dilute!
Note: Store in the -20°C freezer in the box marked "Other Enzymes"

==Media Additives==
===1 M Calcium Chloride (75 mL)===
* 75 mL ddH<sub>2</sub>O
* 11.027 g calcium chloride dihydrate (CaCl<sub>2</sub>·2H<sub>2</sub>O)
Autoclave

===2 M Magnesium Chloride (75 mL)===
* 75 mL ddH<sub>2</sub>O
* 30.4965 g magnesium chloride hexahydrate (MgCl<sub>2</sub>·6H<sub>2</sub>O)
Autoclave

===1 M Sodium Citrate (40 mL)===
* 40 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
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 ddH<sub>2</sub>O
* 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 ddH<sub>2</sub>O
* 7.98 g CuSO<sub>4</sub>
Sterile filter

===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
* 21.915 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 -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