User:Karmella Haynes/Notebook/BioBrick cloning/2012/11/30

(Difference between revisions)

Revision as of 22:51, 30 November 2012

Karmella's BioBrick Cloning

Assembly: CMVTetO2:K (KAH3)/ MV; expression vector for Vi's mammalian cell expression experiment

Assemblies

> Digests (Fermentas FD)

Reagent	Volume	Image:GelImage.jpg 30 μL/lane, 1% agarose; Ladder
DNA (plasmid)	up to 25 μL
10x buffer	3.0
enzyme 1	1.0
enzyme 2	1.0
dH₂O	---
	30 μL --> 37°C/ ~15 min.

> Measure conc.'s of gel purified fragments

> Ligations

@@ Line 42: / Line 42: @@
 | <u>Sample</u> || <u>OD260</u> || <u>260/280</u> || <u>ng/μL</u>
 |-
-| 1. KAH3 (X/P) || --- || --- || ---
+| 1. KAH3 (X/P) || 0.028 || 1.96 || 28.41
 |-
-| 2. MV2 (S/P) || --- || --- || ---
+| 2. MV2 (S/P) || 0.085 || 1.93 || 85.02
 |}
@@ Line 60: / Line 60: @@
 | &nbsp;             || 1    || 2    ||
 |-
-| Insert DNA         || ###  || ---  ||
+| Insert DNA         || 0.3  || ---  ||
 |-
-| Vector DNA         || ###  || ###  ||
+| Vector DNA         || 0.3  || 0.3  ||
 |-
 | 2x lgn buf (Roche) || 5.0  || 5.0  ||
@@ Line 68: / Line 68: @@
 | T4 ligase (NEB)    || 1.0  || 1.0  ||
 |-
-| dH<sub>2</sub>O    || ###  || ###  ||
+| dH<sub>2</sub>O    || 3.4  || 3.7  ||
 |-
 | &nbsp;             || 10 μL || 10 μL ||