1,721
edits
Richard Lab:Restriction Digest: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Richard Lab:Restriction Digest (view source)
Revision as of 15:40, 12 January 2011
, 12 January 2011→Procedure
| Line 17: | Line 17: | ||
# Quickly vortex all ingredients (Buffer, BSA, DNA) before beginning. | # Quickly vortex all ingredients (Buffer, BSA, DNA) before beginning. | ||
# Add the following in a micro-centrifuge tube: | # Add the following in a micro-centrifuge tube: | ||
##5μl of Buffer (usually NEBuffer 2) | ##5μl of Buffer (usually NEBuffer 2); | ||
##1μl of BSA | ##1μl of BSA; | ||
##0.5 picomoles DNA (see [[DNA Quantification]])[[user:Michael A. Speer | Mike]] normally uses 10μL of miniprep or 5μL of purified PCR product | ##0.5 picomoles DNA (see [[DNA Quantification]]);[[user:Michael A. Speer | Mike]] normally uses 10μL of miniprep or 5μL of purified PCR product. | ||
##Water to make 48μl | ##Water to make 48μl. | ||
# Vortex Enzymes and add 1μl (20 units) of each to the tube. | # Vortex Enzymes and add 1μl (20 units) of each to the tube. | ||
# Incubate reaction in a 37°C water bath for at least one hour. | # Incubate reaction in a 37°C water bath for at least one hour. | ||
