Haynes Lab:Notebook/Short Projects/2015/05/28: Difference between revisions
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Line 22: | Line 22: | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Rxn | | bgcolor=#cfcfcf | Rxn | ||
| bgcolor=#cfcfcf | Mix ( | | bgcolor=#cfcfcf | Mix (5x) | ||
|- | |- | ||
| 100 μM Oligo 1 || 1.0 || --- | | 100 μM Oligo 1 || 1.0 || --- | ||
Line 28: | Line 28: | ||
| 100 μM Oligo 2 || 1.0 || --- | | 100 μM Oligo 2 || 1.0 || --- | ||
|- | |- | ||
| 10x T4 Lign buf (NEB) || 1.0 || | | 10x T4 Lign buf (NEB) || 1.0 || 5.0 | ||
|- | |- | ||
| T4 PNK (NEB) || 0.5 || | | T4 PNK (NEB) || 0.5 || 2.5 | ||
|- | |- | ||
| dH<sub>2</sub>O || 6.5 || | | dH<sub>2</sub>O || 6.5 || 32.5 | ||
|- | |- | ||
| || 10.0 | | || 10.0 | ||
Line 59: | Line 59: | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | Rxn | | bgcolor=#cfcfcf | Rxn | ||
| bgcolor=#cfcfcf | Mix ( | | bgcolor=#cfcfcf | Mix (x5) | ||
|- | |- | ||
| 100 ng Vector || 0.5 || | | 100 ng Vector || 0.5 || 2.5 | ||
|- | |- | ||
| 1:250 dsOligo || 2.0 || --- | | 1:250 dsOligo || 2.0 || --- | ||
|- | |- | ||
| 10x FD buf || 2.0 || | | 10x FD buf || 2.0 || 10.0 | ||
|- | |- | ||
| 10 mM DTT || 1.0 || | | 10 mM DTT || 1.0 || 5.0 | ||
|- | |- | ||
| 10 mM ATP || 1.0 || | | 10 mM ATP || 1.0 || 5.0 | ||
|- | |- | ||
| FastDigest BbsI || 1.0 || | | FastDigest BbsI || 1.0 || 5.0 | ||
|- | |- | ||
| T4 DNA ligase || 0.5 || | | T4 DNA ligase || 0.5 || 2.5 | ||
|- | |- | ||
| dH<sub>2</sub>O || 12.0 || | | dH<sub>2</sub>O || 12.0 || 60.0 | ||
|- | |- | ||
| || 20.0 | | || 20.0 |
Revision as of 12:39, 28 May 2015
Haynes Lab - Short Projects | <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> | |||||||||||||||||||||||||||||||||||||||||||||||||
Ben Nyer - 28/05/2015Cloning gRNAs into pSPgRNA for dCas9-HT binding to luc gene
Phosphorylate and anneal oligo pairs
LabNet OptiMax Thermocycler: BbsI Dig/Lig
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