User:Wesley J. Houston/Notebook/MAP/2013/02/08

(Difference between revisions)

Revision as of 01:47, 7 February 2013

Building a Reporter Gene

Assembly: 5xGal4 : [Spacer : HSVtk TATA] : K : AmCyan : AmCyan : NLS : STOP : [polyA2] KAH201/KAH182; AmCyan reporter gene for chromatin proteins.

Assemblies

> Digests (Fermentas FD)

> Measure conc.'s of gel purified fragments

> Ligations (edit after ligation is done)

@@ Line 12: / Line 12: @@
 ----
 '''Assemblies'''
-# KAH201/KAH182: (1) KAH201/(S/P)/203 + (2) KAH182/(X/P)/1675
+# KAH201/KAH182: (1) KAH201/(X/P)/203 + (2) KAH182/(S/P)/1675
 > Digests (Fermentas FD)<br>
@@ Line 22: / Line 22: @@
 | rowspan="9" | [[Image:KAH113012_gel1.jpg‎ |150px|Hover name]]<br>30 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 |-
-| DNA (plasmid) || up to 25 μL
+| DNA (plasmid) || 15 to 25 μL
 |-
 | 10x buffer || 3.0
@@ Line 30: / Line 30: @@
 | enzyme 2 || 1.0
 |-
-| dH<sub>2</sub>O || ---
+| dH<sub>2</sub>O || 0 to 10 μL
 |-
 | &nbsp; || 30 μL --> 37°C/ ~15 min.
@@ Line 41: / Line 41: @@
 | <u>Sample</u> || <u>OD260</u> || <u>260/280</u> || <u>ng/μL</u>
 |-
-| 1. KAH201 (S/P) || TBD || TBD || TBD
+| 1. KAH201 (X/P) || TBD || TBD || TBD
 |-
-| 2. KAH182 (X/P) || TBD || TBD || TBD
+| 2. KAH182 (S/P) || TBD || TBD || TBD
 |}
@@ Line 51: / Line 51: @@
 | <u>Ligation</u> || <font color="blue"><u>Plate results (lig : neg crtl)</u> 11/31/12</font>
 |-
-| 1. KAH201 (S/P)/203, 8 ng + KAH182 (X/P)/1675, 25 ng || <font color="blue">KAH3/MV2 >10:1 (Pick 2)</font>
+| 1. KAH201 (X/P)/203, x ng + KAH182 (S/P)/1675, x ng || <font color="blue">---</font>
 |-
-| 2. KAH182 (X/P)/ 25 ng || zero colonies
+| 2. KAH182 (S/P)/ 25 ng || ---
 |}