Difference between revisions of "User:Madeline Hartman/Notebook/Dr. Hartings Lab/2012/11/06"

From OpenWetWare
Jump to: navigation, search
(Entry title)
(fix raw html notebook nav)
 
(One intermediate revision by one other user not shown)
Line 2: Line 2:
 
|-
 
|-
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
+
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
 
|-
 
|-
 
| colspan="2"|
 
| colspan="2"|
Line 9: Line 9:
 
I ran two PCR reactions with the following content:
 
I ran two PCR reactions with the following content:
  
 +
Tube 1-
 
# 40.6 μL distilled H2O
 
# 40.6 μL distilled H2O
 
# 5.0 μL 10x cloned Pfu buffer
 
# 5.0 μL 10x cloned Pfu buffer
Line 16: Line 17:
 
# 1.0 μL Primer 2 (ng/μL)
 
# 1.0 μL Primer 2 (ng/μL)
 
# 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
 
# 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
 +
 +
and
 +
 +
Tube 2-
 +
# 20.3 μL distilled H2O
 +
# 5.0 μL 10x cloned Pfu buffer
 +
# 0.4 μL dNTPs (25 mM each)
 +
# 1.0 μL DNA template (100 ng/μL)
 +
# 1.0 μL Primer 1 (ng/μL)
 +
# 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
 +
Tube 3-
 +
# 20.3 μL distilled H2O
 +
# 5.0 μL 10x cloned Pfu buffer
 +
# 0.4 μL dNTPs (25 mM each)
 +
# 1.0 μL DNA template (100 ng/μL)
 +
# 1.0 μL Primer 2 (ng/μL)
 +
# 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)
 +
 +
Tubes 2 and 3 were later combined.
 +
 +
Primer 1- t109c forward
 +
 +
Primer 2- t109c reverse
 +
 +
The three tubes were ran for the following cycles:
 +
 +
[[Image:MH 20121106Table of cycles.png|300px]]
 +
 +
 +
After 5 cycles, tubes 2 and 3 were combined, and run the rest of the cycles together.
 +
 +
Tubes were left in thermocycler overnight at 4°C
  
  

Latest revision as of 21:19, 26 September 2017

Owwnotebook icon.png Project name Report.pngMain project page
Next entryResultset next.png

Entry title

I ran two PCR reactions with the following content:

Tube 1-

  1. 40.6 μL distilled H2O
  2. 5.0 μL 10x cloned Pfu buffer
  3. 0.4 μL dNTPs (25 mM each)
  4. 1.0 μL DNA template (100 ng/μL)
  5. 1.0 μL Primer 1 (ng/μL)
  6. 1.0 μL Primer 2 (ng/μL)
  7. 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)

and

Tube 2-

  1. 20.3 μL distilled H2O
  2. 5.0 μL 10x cloned Pfu buffer
  3. 0.4 μL dNTPs (25 mM each)
  4. 1.0 μL DNA template (100 ng/μL)
  5. 1.0 μL Primer 1 (ng/μL)
  6. 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)

Tube 3-

  1. 20.3 μL distilled H2O
  2. 5.0 μL 10x cloned Pfu buffer
  3. 0.4 μL dNTPs (25 mM each)
  4. 1.0 μL DNA template (100 ng/μL)
  5. 1.0 μL Primer 2 (ng/μL)
  6. 1.0 μL Pfu Turbo DNA Polymerase (2.5 U/μL)

Tubes 2 and 3 were later combined.

Primer 1- t109c forward

Primer 2- t109c reverse

The three tubes were ran for the following cycles:

MH 20121106Table of cycles.png


After 5 cycles, tubes 2 and 3 were combined, and run the rest of the cycles together.

Tubes were left in thermocycler overnight at 4°C