Haynes Lab:Notebook/Engineering PC-TFs/2013/01/25

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Autocreate 2013/01/25 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs)
(Summary)
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. ##### -->
-
==Summary==
+
==01/25/13==
-
*  
+
* Type IIS assembly - day 1
 +
 
 +
 
 +
----
 +
 
 +
PCR
 +
* Resuspend new primers to a final concentration of 100 μM
 +
* Make 10 μM dilutions in separate tubes
 +
* All of the reactions are as follows:
 +
 
 +
# template, primer 1, primer 2
 +
# template, primer 1, primer 2
 +
# template, primer 1, primer 2
 +
# template, primer 1, primer 2
 +
 
 +
 
 +
{| {{table}} border="1" cellspacing="3" <!-- PCR rxn. table -->
 +
|- valign="top"
 +
| bgcolor=#cfcfcf | Reagent
 +
| bgcolor=#cfcfcf | Volume
 +
| rowspan="7" | <u>Expected:</u><br>1. pSB1A3 = ~2000<br>2. hPCD = 186<br>3. BL01 = 2520
 +
| rowspan="7" | [[Image:KAH011013_gel1.jpg|200px|Hover name]]<br>10 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 +
|-
 +
| DNA(plasmid) || 0.2 μL
 +
|-
 +
| primer 1 || 1.0
 +
|-
 +
| primer 2 || 1.0
 +
|-
 +
| 2x GoTaq || 25.0
 +
|-
 +
| dH<sub>2</sub>O || 22.8
 +
|-
 +
| &nbsp; || 50.0 μL
 +
|}
 +
 
 +
PCR
 +
* 95°C, 3 min.
 +
* [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
 +
* 72°C, 3 min.
 +
* 4°C, ∞
 +
 

Revision as of 16:38, 25 January 2013

Engineering PC-TFs Main project page
Previous entry      Next entry

01/25/13

  • Type IIS assembly - day 1



PCR

  • Resuspend new primers to a final concentration of 100 μM
  • Make 10 μM dilutions in separate tubes
  • All of the reactions are as follows:
  1. template, primer 1, primer 2
  2. template, primer 1, primer 2
  3. template, primer 1, primer 2
  4. template, primer 1, primer 2


Reagent Volume Expected:
1. pSB1A3 = ~2000
2. hPCD = 186
3. BL01 = 2520
Hover name
10 μL/lane; 1% agarose; Ladder
DNA(plasmid) 0.2 μL
primer 1 1.0
primer 2 1.0
2x GoTaq 25.0
dH2O 22.8
  50.0 μL

PCR

  • 95°C, 3 min.
  • [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
  • 72°C, 3 min.
  • 4°C, ∞



Personal tools