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Recipes

                                                                                                                                                                                                                 

A pinch of salt makes all the difference !!

Preparation of TAE buffer

     20x TAE buffer

• 80mL of 1M Tris solution
• 0.7264g of EDTANa2+ salt
• 2.284 mL glacial acetic acid
• distilled H2O 100mL

Preparation of Ethidium Bromide(EtBr)

• For the preparation of ethidium bromide adds 1 g of ethidium bromide to 100 ml of H2O.
• Stir on a magnetic stirrer for several hours to ensure that the dye has dissolved.
• Wrap the container in aluminum foil or transfer the 10 mg/ml solution to a dark bottle and store at room temperature.

Preparation of alkaline lysis solution I

• 50 mM glucose.
• 25 mM Tris-Cl (pH 8.0).
• 10 mM EDTA (pH 8.0).

Prepare Solution I from standard stocks in batches of approx. 100 ml, sterilize by autoclaving and store at 4°C.

Preparation of alkaline lysis solution II (freshly prepared at room temperature)

• 0.2 N NaOH (freshly diluted from a 10 N stock).
• (100ul SDS + 700ul mq + 200ul NaOH) 1% (w/v) SDS.

Preparation of alkaline lysis solution III

• 5 M potassium acetate, 60.0 ml
• Glacial acetic acid, 11.5 ml
• H2O, 28.5 ml

The resulting solution is 3 M with respect to potassium and 5 M with respect to acetate. Store the solution at 4°C and transfer it to an ice bucket just before use.

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	E-mail: iitd.igem@gmail.com Undergraduate Laboratory Department of Biotechnology and Biochemical Engineering, IIT Delhi