How To Make A Stock Solution

Antibiotics

Ampicillin (forty mL)

For a stock solution of 25 mg/mL:

• ane gAmpicillin
• 40 mL ddH_iiO
• Filter sterilize

Note: Store at iv°C

Chloramphenicol (forty mL)

For a stock solution of 7.five mg/mL:

• 300 mgChloramphenicol
• forty mL 50%Ethanol

Notation: Do not filter sterilize—the ethanol may deliquesce the filter
Note: Shop at 4°C

Erythromycin (20 mL)

For a stock solution of 10 mg/mL:

• 200 mgErythromycin
• 20 mL 100% ethanol (in smoke hood)
• Donot filter sterilize—the ethanol may deliquesce the filter

NOTE: Store at –20°C

Gentamicin (xx mL)

For a stock solution of 7.5 mg/mL:

• 150 mgGentamicin
• twenty mL ddH₂O
• Filter sterilize

Annotation: Store at 4°C

Hygromycin (40 mL)

For a stock solution of v mg/mL:

• 200 mgHygromycin
• twoscore mL ddH_iiO
• Filter sterilize

Annotation: Store at 4°C

Kanamycin (40 mL)

For a stock solution of seven.5 mg/mL:

• 300 mgKanamycin
• 40 mL ddH_twoO
• Filter sterilize

NOTE: Shop at 4°C

Nalidixic acid (30 mL)

For a stock solution of thirty mg/mL:

• 900 mgNalidixic Acid
• 360 mg NaOH
• thirty mL ddH_twoO
• Filter sterilize

NOTE: Store at 4°C

Neomycin (twoscore mL)

For a stock solution of 25 mg/mL:

• 1 gNeomycin
• twoscore mL ddH₂O
• Filter sterilize

Note: Store at 4°C

Phosphomycin

For a stock solution of 25 mg/mL:

• one gPhosphomycin
• xl mL ddH_iiO
• Filter sterilize

Note: Store at 4°C

Rifampicin (20 mL)

For a stock solution of l mg/mL:

• 1 gRifampicin
• 20 mL North,Due north-dimethylformamide (DMF; in fume hood)

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

Spectinomycin (xl mL)

For a stock solution of twenty mg/mL:

• 800 mgSpectinomycin
• 40 mL ddH_twoO
• Filter sterilize

NOTE: Store at four°C

Streptomycin (40 mL)

For a stock solution of fifty mg/mL:

• ii kStreptomycin
• forty mL ddH₂O
• Filter sterilize

NOTE: Shop at four°C

Tetracycline (xl mL)

For a stock solution of 2.5 mg/mL:

• 100 mgTetracycline
• 40 mL 50%Ethanol

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

Triclosan (twoscore mL)

For a stock solution of 10 mg/mL:

• 400 mgTriclosan
• forty mL 100%Ethanol

Annotation: Practise 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 mgTrimethoprim
• 40 mL 100%DMSO

Note: Practice not filter sterilize—the DMSO may dissolve the filter
Note: Store at 4°C

Antibiotics Tabular array

Deoxyribonucleic acid

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 μldGTP
  • x μldATP
  • 10 μldTTP
  • ten μldCTP
• Manipulate 100-μL aliquots into 1.7-mL tubes

Note: Store in the –xx°C freezer in the box marked "PCR Supplies"

10X

Taq

buffer

• Characterization tubes: 10XTaq buffer
• Thaw and mix (stir) "Thermo political leader buffer" (Neb; in freezer)
• Aliquot 375 μL of "Thermo political leader buffer" into tubes—donot dilute!

Notation: Store in the –20°C freezer in the box marked "PCR Supplies"

10X

Pfx

buffer

• Label tubes: 10XPfx buffer
• Thaw and mix (stir) "Pfx50 10X PCR Buffer" (NEB; in freezer)
• Aliquot 375 μL of "Pfx50 10X PCR Buffer" into tubes—donot dilute!

Note: Store in the –20°C freezer in the box marked "PCR Supplies"

5X

Q5

buffer

• Characterization tubes: 5XQ5 buffer
• Thaw and mix (stir) "Q5 5X PCR Buffer" (NEB; in freezer)
• Aliquot 100 μL of "Q5 5X PCR Buffer" into tubes—practisenon dilute!

Note: Shop in the –xx°C freezer in the box marked "PCR Supplies"

8X Dna loading

• forty%glycerol
• 1/50 dilution of2% bromophenol blue

DNA ladder

• 140 μL ddH_ii0
• 40 μL8X Dna loading  buffer
• twenty μL 1 kb ladder from Bill
• 2 μL0.25 M EDTA

Notation: use four μL per lane; 3kb ring = 50 ng

Ethidium Bromide (1 mL)

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

Notation: use viii μL per 50 mL gel
Warning: Always vesture gloves when treatment ethidium bromide

Ethidium Bromide (10 mL)

• 10 mL ddH₂O
• x mg ethidium bromide

Annotation: use viii μL per 50 mL gel
WARNING: Always wear gloves when handling ethidium bromide

Media Additives

1 Grand Calcium Chloride (75 mL)

• 75 mL ddH_twoO
• 11.027 g calcium chloride dihydrate (CaCl₂·2H_iiO)

Autoclave

ii M Magnesium Chloride (75 mL)

• 75 mL ddH₂O
• thirty.4965 g magnesium chloride hexahydrate (MgCl₂·6H₂O)

Autoclave

1 M Sodium Citrate (40 mL)

• 35 mL ddH₂O
• 11.76 g sodium citrate tribasic dihydrate (Na₃C₆H_vO₇·2H_twoO)
• pH to 5.7 with HCl (~1 mL)
• QS to forty mL with ddH_twoO

Sterile filter

2% X-gal

• x mL N,N-dimethylformamide (DMF)(in fume hood)
• 0.2 g v-bromo-iv-chloro-3-indolyl- beta-D-galactopyranoside (Ten-gal)(in freezer)

Store at –twenty°C

0.2 Yard IPTG

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

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

i M Copper (Two) sulfate (l mL)

• 50 mL ddH₂O
• 7.98 g CuSO₄

Sterile filter

10%

SDS

(75 mL)

• 75 mL ddH₂O
• vii.5 g Sodium dodecyl sulfate (SDS )

Do non sterilize

20%

SDS

(75 mL)

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

Practise not sterilize

30% Acrylamide (75 mL)

• 75 mL ddH₂O
• 22.five g acrylamide

twoscore% Acrylamide (75 mL)

• 75 mL ddH₂O
• 30 thousand acrylamide

ten% Ammonium persulfate (75 mL)

• 75 mL ddH₂O
• 7.five k ammonium persulfate (APS)

five One thousand NaCl (75 mL)

• 75 mL ddH_iiO
• 21.92 one thousand NaCl

Autoclave

80% Glycerol (75 mL)

• 15 mL ddH₂O
• sixty mL glycerol

Microwave for 30-2d intervals on high until dissolved
Autoclave

i M DTT (5 mL)

• five 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)
• ten mL sulfuric acid (H_iiSo_four (in fume hood—be conscientious!)
• 200 mg orcinol

Source: https://mmbio.byu.edu/griffitts-lab/stock-solution

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