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

(Difference between revisions)

Revision as of 14:10, 4 April 2012

@@ Line 16: / Line 16: @@
 <!-- Precede finished items with a checkmark &#x2713; -->
 * &#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__