User:Karmella Haynes/Notebook/BioBrick cloning/2015/05/28: Difference between revisions
From OpenWetWare
Line 16: | Line 16: | ||
* Assemblies | * Assemblies | ||
# | # DT001_pUC57: hPCD(E/S)/186 + PLflex_pUC57(E/X)/2763 | ||
# | # DT002_pUC57: hPCD(E/S)/186 + PLflex4_pUC57(E/X)/2930 | ||
# | # DT003_pUC57: hPCD(E/S)/186 + PLrigid_pUC57(E/X)/2763 | ||
# | # DT004_pUC57: hPCD(E/S)/186 + PLrigid4_pUC57(E/X)/2931 | ||
* Digests (Fermentas FD) | * Digests (Fermentas FD) | ||
{| {{table}} cellspacing="3" <!-- Digest rxn. table --> | {| {{table}} cellspacing="3" <!-- Digest rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | | | bgcolor=#cfcfcf | Rxn1 | ||
| rowspan="7" | < | | bgcolor=#cfcfcf | Rxn2-5 | ||
| rowspan="7" | Expected bands:<br>1. hPCD(E/S) = 186<br>2. PLflex_pUC57(E/X) = 2763<br>3. PLflex4_pUC57(E/X) = 2930<br>4. PLrigid_pUC57(E/X) = 2763<br>5. PLrigid4_pUC57(E/X) = 2931 | |||
| rowspan="7" | [[Image:somegel.jpg|270px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] | |||
|- | |- | ||
| DNA (plasmid) || | | DNA (plasmid) || 25.0 || 15.0 | ||
|- | |- | ||
| 10x buffer || 3.0 | | 10x buffer || 3.0 || 3.0 | ||
|- | |- | ||
| enzyme 1 || 1.0 | | enzyme 1 || 1.0 || 1.0 | ||
|- | |- | ||
| enzyme 2 || 1.0 | | enzyme 2 || 1.0 || 1.0 | ||
|- | |- | ||
| dH<sub>2</sub>O || --- | | dH<sub>2</sub>O || --- || 10.0 | ||
|- | |- | ||
| || 30 μL | | || 30 μL | ||
|} | |} | ||
--> 37°C/ ~30 min. | |||
* Gel purification | |||
** Sigma GenElute Gel purification kit | |||
** Elute & back-elute w/ 25 μL elution sln. | |||
Line 50: | Line 59: | ||
| Sample || OD260 || 260/280 || ng/μL | | Sample || OD260 || 260/280 || ng/μL | ||
|- | |- | ||
| 1. | | 1. hPCD(E/S) || --- || --- || --- | ||
|- | |- | ||
| 2. | | 2. PLflex_pUC57(E/X) || --- || --- || --- | ||
|} | |} | ||
Revision as of 17:26, 28 May 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> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||
05/28/15
David Tze - assemblies: linkers + hPCD
--> 37°C/ ~30 min.
|