Oneill Lab:Enzymes

Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology

Lab Stock Enzymes
Preparation of lab stock enzymes and buffers

10x EcoRI Buffer

 * 1) Combine
 * 2) *8mL 2.5M Tris
 * 3) *2mL 5M  NaCl
 * 4) *1mL 1M  MgCl2
 * 5) *50&mu;L TritonX-100
 * 6) Adjust pH to 7.5 with 12M HCl
 * 7) Add water to 20mL

Proteinase K
Makes 0.5mL at 20mg/ml
 * 10&mu;L 1M CaCl2
 * 5&mu;L 1M Tris pH 7.5
 * 250&mu;L Glycerol
 * Pipette with cut tip
 * Use under flame
 * 10mg ProK
 * Stored in 4&deg;C dessicator
 * 235&mu;L H2O

Aliquot and store at -20&deg;C

Nuclease Solution
Used in lambda extractions prior to capsid degradation.
 * DNaseI 10mg
 * RNaseA 10mg
 * 1M Tris 10&mu;l
 * 5M NaCl 30&mu;l
 * 1M MgCl2 1&mu;l
 * Water 950&mu;l

Use at concentration of 1mg/ml

Links and Contacts
Seth Kasowitz [mailto:seth.kasowitz@uconn.edu]

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