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

From OpenWetWare
Jump to navigationJump to search
Line 18: Line 18:


# pSB1A3, gg0001, gg0002
# pSB1A3, gg0001, gg0002
# template, primer 1, primer 2
# hPCD, gg0003, gg0004
# template, primer 1, primer 2
# BL02, gg0005, gg0007
# template, primer 1, primer 2
# BL03, gg0005, gg0008
# BL04, gg0005, gg0009
# fshPCD, gg0010, gg0012




Revision as of 22:34, 27 January 2013

Engineering PC-TFs <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>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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. pSB1A3, gg0001, gg0002
  2. hPCD, gg0003, gg0004
  3. BL02, gg0005, gg0007
  4. BL03, gg0005, gg0008
  5. BL04, gg0005, gg0009
  6. fshPCD, gg0010, gg0012


Reagent Volume Expected:
1. pSB1A3 = ~2000
2. hPCD = 186
3. BL02 = 1431
4. BL03 = 1731
5. BL04 = 1686
6.fshPCD = 186
 PCR results
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
  • 95°C, 3 min.
  • [95°C, 30 sec.; 57°C, 30 sec.; 72°C, 1 min.] x35
  • 72°C, 3 min.
  • 4°C, ∞


Purification

  • Clean with Zymo DNA c&c; elute w/ 20 μL dH2O
  • Measure [DNA]
Sample OD 260 260/280 ng/μL
1. pSB1A3 ### ### ###
2. ### ### ### ###
3. ### ### ### ###



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Engineering_PC-TFs/2013/01/25&oldid=670922"