Haynes Lab:Notebook/Engineering PC-TFs/2012/10/26: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(11 intermediate revisions by the same user not shown) | |||
Line 9: | Line 9: | ||
{|border="1" cellpadding="5" cellspacing="0" align="left" | {|border="1" cellpadding="5" cellspacing="0" align="left" | ||
|- | |- | ||
! scope="col" style="background:#efefef;" | | ! scope="col" style="background:#efefef;" | Assembly # | ||
! scope="col" style="background:#efefef;" | Concentration | ! scope="col" style="background:#efefef;" | Concentration | ||
Line 36: | Line 36: | ||
<br> | <br> | ||
<br> | <br> | ||
<br> | |||
<br> | |||
*<u>Gibson Assembly </u> | |||
Assembly 1 contains PCR rxns 1,2, and 3. Assembly 2 contains PCR rxns 4,5. | |||
{|border="1" cellpadding="5" cellspacing="0" align="left" | |||
|- | |||
! scope="col" style="background:#efefef;" | | |||
! scope="col" style="background:#efefef;" | 1 | |||
! scope="col" style="background:#efefef;" | 2 | |||
|- | |||
|Gibson Rxn mix | |||
|15 μL | |||
|15 μL | |||
|- | |||
|DNA fragments | |||
|5 μL | |||
|5 μL | |||
|} | |||
<br> | <br> | ||
<br> | <br> | ||
Line 41: | Line 63: | ||
<br> | <br> | ||
<br> | <br> | ||
{|border="1" cellpadding="5" cellspacing="0" align="left" | |||
|- | |||
! scope="col" style="background:#efefef;" | <i>DNA fragments</i> | |||
! scope="col" style="background:#efefef;" | 1 | |||
! scope="col" style="background:#efefef;" | 2 | |||
|- | |||
|#1 | |||
|1.0 μL | |||
| | |||
|- | |||
|#2 | |||
|1.0 μL | |||
| | |||
|- | |||
|#3 | |||
|1.0 μL | |||
| | |||
|- | |||
|#4 | |||
| | |||
|1.0 μL | |||
|- | |||
|#5 | |||
| | |||
|1.0 μL | |||
|- | |||
|Vector | |||
|1.0 μL | |||
|1.0 μL | |||
|- | |||
|dH2O | |||
|1.0 μL | |||
|2.0 μL | |||
|- | |||
|<i>Total</i> | |||
|5.0 μL | |||
|5.0 μL | |||
|} | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
<br> | |||
*Thaw DHSα Turbo cells (50 μL per transformation) | |||
*Add 20 μL of Gibson reaction to 50μL DHSα Turbo. | |||
**Assembly 1 | |||
**Assembly 2 | |||
**Negative Control (DH2O) | |||
*Incubate on ice for five minutes. | |||
*Plate 70 μL transformation on warm Amp plates. | |||
Revision as of 13:39, 30 October 2012
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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> | ||||||||||||||||||||||||||||||||||||||||||||||||
Summary
Assembly 1 contains PCR rxns 1,2, and 3. Assembly 2 contains PCR rxns 4,5.
|