User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
Line 16: Line 16:
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* &#x2713; Assemblies: BD111, BD112, BD113
* &#x2713; Assemblies: BD111, BD112, BD113
-
----
 
-
'''Assemblies'''
 
-
# BD111: KAH109/(X/P)/1123 + BBa_B0030/(S/P)/15
 
-
# BD112: BD111/(X/P)/1138 + BBa_I712074/(S/P)46
 
-
# BD113: BD111/(X/P)/1138 + BBa_I719005/(S/P)23
 
-
 
-
 
-
> Digests (Fermentas FD)<br>
 
-
 
-
{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
 
-
|-valign="top"
 
-
| <u>Reagent</u> || <u>Volume</u> || &nbsp;
 
-
| rowspan="9" |
 
-
|-
 
-
| DNA (plasmid) || up to 20 μL
 
-
|-
 
-
| 10x buffer || 3.0
 
-
|-
 
-
| enzyme 1 || 1.0
 
-
|-
 
-
| enzyme 2 || 1.0
 
-
|-
 
-
| dH<sub>2</sub>O || ---
 
-
|-
 
-
| &nbsp; || 30 μL --> 37°C/ ~30 min.
 
-
|}
 
-
{| class="wikitable" width=400px
 
-
| Plasmid DNA || 15.0 μl*
 
-
|-
 
-
| Fermentas FastDigest enzyme 1 || 1.0 μl
 
-
|-
 
-
| Fermentas FastDigest enzyme 2 || 1.0 μl
 
-
|-
 
-
| 10x FastDigest buffer + green loading dye || 3.0 μl
 
-
|-
 
-
| dH<sub>2</sub>O || 10.0 μl
 
-
|-
 
-
| &nbsp; || 30.0 μl total
 
-
|-
 
-
| colspan="2" | *For low yield DNA, use up to 25 μL; decrease dH<sub>2</sub>O accordingly.<br>Mix the reaction(s) thoroughly by flicking the tube.<br>Incubate at 37°C for 10 minutes.
 
-
|}
 
-
__NOTOC__
 

Revision as of 13:10, 4 April 2012

Search this Project

Customize your entry pages

04/04/11

  • ✓ Assemblies: BD111, BD112, BD113
Personal tools