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Microsoft word - drug plate.doc

YES-HU plate
Preparation of HU (Hydroxyurea) Stock Solution; 1 M

Hydroxyurea … MW: 76.05 @ 4 C
Dissolve 0.76 g in 10 ml water. Aliquot in 1.5 ml tubes. Keep –20 C.
Preparation of YES-HU plates
Make 1 L of YES with 20g agar in a 2 L plastic flask with a big stir bar. Autoclave 45
min. Stir slowly until it gets cool enough to add the drug, like you can touch it with bare
hands (~60˚C). Add HU Stock Solution and mix thoroughly. Then pour into Petri dishes
before it starts solidify. 1 L makes 2 sleeves (40) of plates. Let plates dry for 2 days, put
them back in sleeves and store at 4˚C room. The drug in plate last for a year.
2 mM HU plate: 2.0 ml / 1 L
5 mM HU plate: 5.0 ml / 1 L

YES-CPT plate
Preparation of CPT (Camptothecin) Stock Solution; 10 mM
Camptothecin … MW: 348.36 @ 4 C
Dissolve 34.8 mg CPT in 10 ml DMSO. Aliquot in 1.5 ml tubes. Keep –20 C.
Preparation of YES-CPT plates
Make 1 L of YES with 20g agar in a 2 L plastic flask with a big stir bar. Autoclave 45
min. Stir slowly until it gets cool enough to add the drug, like you can touch it with bare
hands (~60˚C). Add CPT Stock Solution and mix thoroughly. Then pour into Petri dishes
before it starts solidify. 1 L makes 2 sleeves (40) of plates. Let plates dry for 2 days, put
them back in sleeves and store at 4˚C room. The drug in plate last for a year.
2 µM CPT plate: 0.2 ml / 1 L
5 µM CPT plate: 0.5 ml / 1 L


YES-G418 plate
Preparation of G418 Stock Solution; 50 mg/ml
Geneticin concentration… 733 µg/mg (Check the label. Each lot is different.)
Dissolve 2.046 g into 30 ml of 1M Tris-HCl (pH8.0), and filtrate with a 0.22 µm disk
filter on a 50 ml syringe.
Preparation of YES-G418 plate
Make 1 L of YES with 20g agar in a 2 L plastic flask with a big stir bar. Autoclave 45
min. Stir slowly until it gets cool enough to add the drug, like you can touch it with bare
hands (~60˚C). Add 3 ml G418 Stock Solution and mix thoroughly. Then pour into Petri
dishes before it starts solidify. 1 L makes 2 sleeves (40) of plates. Let plates dry for 2
days, put them back in sleeves and store at 4˚C room. The drug in plate last for a year.
150 mg/l G418 plate: 3 ml / 1 L for 40 plates (2 sleeves)
YES-NAT plate
Preparation of clonNAT Stock Solution; 200 mg/ml (8000x)
clonNAT (Nourseothricin Sulfate) [Werner BioAgents]… powder @4˚C
Dissolve 200 mg into 1 ml of sterilized H O, and filtrate with 0.22 µm filter.
Aliquot 100 µl in 1.5 ml tube, and store at -20˚C.
Preparation of YES-clonNAT plate
Make 500 ml of YES with 20g agar in a 1 L flask with a big stir bar. Autoclave 45 min.
Stir slowly until it gets cool enough to add the drug, like you can touch it with bare hands
(~60˚C). Add clonNAT Stock Solution and mix thoroughly. Then pour into Petri dishes
before it starts solidify. 500 ml makes 1 sleeve (20) of plates. Let plates dry for 2 days,
put them back in sleeves and store at 4˚C room. The drug in plate last for I don’t know.
25 µg/l NAT plate: 125 µl / 500 ml for 20 plates
YES-HYG plate
Preparation of Hygromycin B Stock Solution; 50 mg/ml in PBS (500x)
Hygromycin B [Roche]… 1 g (in 20 ml PBS solution, sterile) @4˚C
Aliquot in 1.5 ml tubes, and store at 4˚C.
Preparation of YES-HYG plate
Make 1 L of YES with 20g agar in a 2 L plastic flask with a big stir bar. Autoclave 45
min. Stir slowly until it gets cool enough to add the drug, like you can touch it with bare
hands (~60˚C). Add 3 ml Hygromycin B Stock Solution and mix thoroughly. Then pour
into Petri dishes before it starts solidify. 1 L makes 2 sleeves (40) of plates. Let plates dry
for 2 days, put them back in sleeves and store at 4˚C room. The drug in plate last for 6
months.
100 mg/l HYG plate: 2 ml / 1 L for 40 plates

Source: http://eishinoguchi.com/drugplate.pdf

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