Melbourne/Diagnostic Digest
From 2007.igem.org
(Difference between revisions)
| Line 1: | Line 1: | ||
| - | ==For 20uL reation volume== | + | ===For 20uL reation volume=== |
| - | ===Reaction Mixture=== | + | ====Reaction Mixture==== |
*0.5uL Enzyme 1 | *0.5uL Enzyme 1 | ||
*0.5uL Enzyme 2 | *0.5uL Enzyme 2 | ||
| Line 10: | Line 10: | ||
**If performing the same digest for multiple DNA samples make up mastermix of buffer enzyme soultion and make 15uL aliquots to which to add the DNA | **If performing the same digest for multiple DNA samples make up mastermix of buffer enzyme soultion and make 15uL aliquots to which to add the DNA | ||
**Volume DNA may vary depending on concentration, vary water volume to achieve 20uL reaction volume. | **Volume DNA may vary depending on concentration, vary water volume to achieve 20uL reaction volume. | ||
| + | |||
#Incubate for 1-3 hrs at 37 degrees. | #Incubate for 1-3 hrs at 37 degrees. | ||
#Stop reaction with addition of 5uL 6x DNA loading dye. | #Stop reaction with addition of 5uL 6x DNA loading dye. | ||
#Can be stored at -20 or run on gel immediately. | #Can be stored at -20 or run on gel immediately. | ||
Revision as of 06:23, 8 July 2007
For 20uL reation volume
Reaction Mixture
- 0.5uL Enzyme 1
- 0.5uL Enzyme 2
- Add enzymes last
- 2uL appropriate 10x buffer (see table on fridge)
- 2uL 10x BSA
- 10uL MilliQ
- 5uL DNA
- If performing the same digest for multiple DNA samples make up mastermix of buffer enzyme soultion and make 15uL aliquots to which to add the DNA
- Volume DNA may vary depending on concentration, vary water volume to achieve 20uL reaction volume.
- Incubate for 1-3 hrs at 37 degrees.
- Stop reaction with addition of 5uL 6x DNA loading dye.
- Can be stored at -20 or run on gel immediately.
