User:Karmella Haynes/Notebook/BioBrick cloning/2015/03/26: Difference between revisions
From OpenWetWare
Line 6: | Line 6: | ||
| colspan="2"| | | colspan="2"| | ||
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit above this line unless you know what you are doing. ##### --> | ||
== | ==03/26/15== | ||
<!-- Precede finished items with a checkmark ✓ --> | <!-- Precede finished items with a checkmark ✓ --> | ||
* '''PEG precipitation - optimization''' | * '''PEG precipitation - optimization''' | ||
Line 14: | Line 14: | ||
---- | ---- | ||
'''PEG precipitation - optimization'''<br> | '''PEG precipitation - optimization'''<br> | ||
* Make PEG | * See OWW protocol - [http://openwetware.org/wiki/Protocol_Size_selective_DNA_precipitation_by_PEG/MgCl2 Size selective DNA precipitation] | ||
* In this procedure, PEG is diluted 3-fold in the final DNA mixture | |||
* Try these final PEG concentrations: 10%, 6%, 5%, 4%, 3%, 2% | |||
'''Make PEG solutions''' | |||
* First. make 30% PEG: 0.3 g PEG 8000 + 10 mL 30 mM MgCL2 | |||
* Use this to make other solutions | |||
{| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | {| {{table}} border="1" cellspacing="3" <!-- Digest check rxn. table --> | ||
|- valign="top" | |- valign="top" | ||
| bgcolor=#cfcfcf | Reagent | | bgcolor=#cfcfcf | Reagent | ||
| bgcolor=#cfcfcf | | | bgcolor=#cfcfcf | 18% PEG | ||
| bgcolor=#cfcfcf | 15% PEG | |||
| bgcolor=#cfcfcf | 12% PEG | |||
| bgcolor=#cfcfcf | 9% PEG | |||
| bgcolor=#cfcfcf | 6% PEG | |||
| rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br> | | rowspan="7" | <u>Expected:</u><br>1. BB 1 = size<br>2. BB2 = size<br> | ||
| rowspan="7" | <!-- [[Image:GelImage.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | | rowspan="7" | <!-- [[Image:GelImage.jpg|400px|Hover name]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder] --> | ||
|- | |- | ||
| | | 30% PEG || | ||
|- | |- | ||
| | | 30 mM MgCl2 || | ||
|- | |- | ||
| dH<sub>2</sub>O || 9.5 | | dH<sub>2</sub>O || 9.5 | ||
|- | |- | ||
| || | | || 10 mL | ||
|} | |} | ||
Revision as of 11:08, 26 March 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> | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
03/26/15
PEG precipitation - optimization
Assemblies
Oligo annealing
|