Haynes Lab:Notebook/Engineering PC-TFs/2013/10/21: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Autocreate 2013/10/21 Entry for Haynes_Lab:Notebook/Engineering_PC-TFs) |
|||
(4 intermediate revisions by the same user 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. ##### --> | ||
== | ==October 21, 2013== | ||
* | <big>''Miniprep liquid cultures''</big> | ||
*Add 100 μL of 7X Lysis Buffer (Blue) to 600 μL of E.coli culture in a 1.5 ml microcentrifuge tube. Mix by inverting tube 4-6times. | |||
*Add 350 μL of cold Neutralization Buffer (Yellow), mix thoroughly. Neutralization is complete when sample becomes yellow and precipitate has formed. | |||
*Centrifuge at 11,000-16,000 g for two minutes. | |||
*Transfer the supernatent into the Zymo-Spin Column. Place column in 2 ml collection tube and centrifuge for 15 seconds at top speed. Discard flow-through. | |||
*Add 200μL of Endo-Wash Buffer to the column. Centrifuge for 15 seconds at top speed. | |||
*Add 400μL of Zyppy Wash Buffer to the column. Centrifuge at top speed for 30 seconds. | |||
*Transfer column to 1.5 microcentrifuge tube and add 50μL of Zyppy Elution Buffer directly to column. Centrifuge for 1 minute at top speed to elute DNA. | |||
<big>''Measure concentration of each fragment''</big>: | |||
* Load Gen 5 2.0 software. Go to the Task Manager-Take 3 Application | |||
* Go to Nucleic Acid Quantification. | |||
* Pipette 2 µL of each DNA sequence into Take 3 plate. | |||
* Place plate in EPOCH plate reader. | |||
{| {{table}} | |||
| align="center" style="background:#efefef;"| '''Part''' | |||
| align="center" style="background:#efefef;"|'''Concentration''' | |||
|- | |||
| KAH126 (1)|| 22.239 ng/μL | |||
|- | |||
| KAH126 (2)||57.672 ng/μL | |||
|- | |||
| CMV+MV9 (1)||11.323 ng/μL | |||
|- | |||
| CMV+MV9 (2)||1.92 ng/μL | |||
|- | |||
| CMV+MV9 (3)||27.394 ng/μL | |||
|} | |||
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> | <!-- ##### DO NOT edit below this line unless you know what you are doing. ##### --> |
Revision as of 14:13, 25 October 2013
<html><img src="/images/9/94/Report.png" border="0" /></html> Main project page <html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | |||||||||||||
October 21, 2013Miniprep liquid cultures
|