User:Karmella Haynes/Notebook/BioBrick cloning/2015/04/16: Difference between revisions
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LabNet OptiMax Thermocycler | LabNet OptiMax Thermocycler: AnOlig RD | ||
* 37°C, 30 min | * 37°C, 30 min | ||
* 95°C, 5 min | * 95°C, 5 min | ||
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| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Rxn | | bgcolor=#cfcfcf | Rxn | ||
| bgcolor=#cfcfcf | Mix (x7) | |||
|- | |- | ||
| 100 ng Vector || 0.5 | | 100 ng Vector || 0.5 || 3.5 | ||
|- | |- | ||
| 1:250 dsOligo || 2.0 | | 1:250 dsOligo || 2.0 || --- | ||
|- | |- | ||
| 10x FD buf (NEB) || 2.0 | | 10x FD buf (NEB) || 2.0 || 14.0 | ||
|- | |- | ||
| 10 mM DTT || 1.0 | | 10 mM DTT || 1.0 || 7.0 | ||
|- | |- | ||
| 10 mM ATP || 1.0 | | 10 mM ATP || 1.0 || 7.0 | ||
|- | |- | ||
| FastDigest BbsI || 1.0 | | FastDigest BbsI || 1.0 || 7.0 | ||
|- | |- | ||
| T7 DNA ligase || 0.5 | | T7 DNA ligase || 0.5 || 3.5 | ||
|- | |- | ||
| dH<sub>2</sub>O || | | dH<sub>2</sub>O || 12.0 || 84.0 | ||
|- | |- | ||
| || 20.0 | | || 20.0 | ||
|} | |} | ||
LabNet OptiMax Thermocycler: BbsI Dig/Lig | |||
* 6x [37°C, 5 min; 23°C, 5 min] | * 6x [37°C, 5 min; 23°C, 5 min] | ||
* 4°C, ∞ | * 4°C, ∞ |
Revision as of 17:49, 16 April 2015
Karmella's BioBrick Cloning | <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
04/16/15
LCR Development
Measure conc.
Program: Phusion (block B)
Conclusion
gRNA cloning - Rene
Phosphorylate and anneal oligo pairs
LabNet OptiMax Thermocycler: BbsI Dig/Lig
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