water

250 mg IPTG (Sigma I-5502)

ddH₂O to 10 ml (aliquot and store at -20℃)

1 M isocitrate (sodium salt-dihydrate):

29.41 g Na3isocitrate-2H₂O (Sigma C-7254)

ddH₂O to 100 ml

10x Kinase buffer: 500 mM Tris-HCl, pH 7.6, 100 mM MgCl2, and 100 mM DTT in

sterile double distilled water.

5 ml 1 M Tris-HCl, pH 7.6

1 ml 1 M MgCl2

1 ml 1 M DTT

sddH₂O to 10 ml (store in 25 ml aliquots at -20deg.C).

Kanamycin sulfate (Kan): Stock of 5 mg/ml in sterile double distilled water (sddH₂O).

0.5 g Kanamycin (Boehringer Mannheim 106 801)

sddH₂O to 100 ml (Add to media for final conc. 20 ug/ml)

1M KCl (potassium chloride):

7.5 g KCl

ddH₂O to 100 ml

10x Ligation buffer: 50 mM Tris-HCl, pH 7.6, 10 mM MgCl2, 10 mM DTT, 10 mM rATP, and 250 ug/ml bovine serum albumin (BSA) in sterile double distilled water.

5 ml 1 M Tris-HCl, pH 7.6

1 ml 1 M MgCl2

1 ml 1 M DTT

1 ml 100 mM rATP

2.5 mg BSA

sddH₂O to 10 ml (store in 25 ml aliquots at -20deg.C)

Loading dye: 0.3% xylene cyanole FF, 0.3% bromophenol blue, 10 mM EDTA in deionized formamide.

3 g xylene cyanole FF

3 g bromophenol blue

0.2 ml 0.5 M EDTA

deionized formamide to 10 ml

Lysozyme solution: 50 mM Tris-HCl, pH 8.0, 10 mM EDTA, and 5 mg/ml lysozyme in sterile double distilled water.

5 ml 1 M Tris-HCl, pH 8.0

2 ml 0.5 M EDTA

0.5 g lysozyme (Sigma L-6876)

sddH₂O to 100 ml (make fresh)

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