User:Karmella Haynes/Notebook/BioBrick cloning/2012/12/03

From OpenWetWare

< User:Karmella Haynes | Notebook | BioBrick cloning | 2012 | 12(Difference between revisions)
Jump to: navigation, search
(mm/dd/yy)
Current revision (19:13, 11 December 2012) (view source)
(12/03/12)
 
(2 intermediate revisions not shown.)
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. ##### -->
-
==mm/dd/yy==
+
==12/03/12==
<!-- Precede finished items with a checkmark &#x2713; -->
<!-- Precede finished items with a checkmark &#x2713; -->
* Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)
* Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)
Line 29: Line 29:
{| {{table}} cellspacing="3" <!-- Ligations table -->
{| {{table}} cellspacing="3" <!-- Ligations table -->
|- bgcolor=#cfcfcf
|- bgcolor=#cfcfcf
-
| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> mm/dd/yy</font>
+
| Ligation || <font color="blue"><u>Plate results (lig : neg crtl)</u> 12/04/12</font>
|-
|-
-
| 1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng || <font color="blue">new BioBrick #:1 (Pick #)</font>
+
| 1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng || <font color="red">KAH111/C-His 0 colonies</font>
|-
|-
-
| 2. C-His(E/P)/ 25 ng || &nbsp;
+
| 2. C-His(E/P)/ 25 ng || 0 colonies
|}
|}
{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
{| {{table}} cellspacing="3" <!-- Ligation rxn table -->
-
| &nbsp;            || 1    || 2    ||
+
| &nbsp;            || 1    || 2     
|-
|-
-
| Insert DNA        || 1.0  || ---  ||
+
| Insert DNA        || 1.0  || ---   
|-
|-
-
| Vector DNA        || 0.5  || 0.5  ||
+
| Vector DNA        || 0.5  || 0.5   
|-
|-
-
| 2x lgn buf (Roche) || 5.0  || 5.0  ||
+
| 2x lgn buf (Roche) || 5.0  || 5.0   
|-
|-
-
| T4 ligase (NEB)    || 1.0  || 1.0  ||
+
| T4 ligase (NEB)    || 1.0  || 1.0   
|-
|-
-
| dH<sub>2</sub>O    || 2.5  || 3.5  ||
+
| dH<sub>2</sub>O    || 2.5  || 3.5   
|-
|-
-
| &nbsp;            || 10 μL || 10 μL ||
+
| &nbsp;            || 10 μL || 10 μL  
|}
|}

Current revision

Karmella's BioBrick Cloning Main project page
Previous entry      Next entry

12/03/12

  • Assembly: KAH111 + C-His expression vector (trouble shooting for Behzad)



Assemblies

  1. KAH111/C-His: KAH111/(E/P)/1123 + C-His/(E/P)/3600


  • Digests (Fermentas FD)
    • Already done by Behzad


  • Measure conc.'s
    • Behzad: KAH111 = 15 ng/μL
    • Behzad: C-His = 57 ng/μL


  • Ligations
Ligation Plate results (lig : neg crtl) 12/04/12
1. KAH111(E/P)/1123, 16 ng + C-His(E/P)/3600, 25 ng KAH111/C-His 0 colonies
2. C-His(E/P)/ 25 ng 0 colonies
  1 2
Insert DNA 1.0 ---
Vector DNA 0.5 0.5
2x lgn buf (Roche) 5.0 5.0
T4 ligase (NEB) 1.0 1.0
dH2O 2.5 3.5
  10 μL 10 μL



Personal tools