Difference between revisions of "Haynes Lab:Notebook/David Barclay Undergrad Training/Plasmid Transformation/2013/10/03"

From OpenWetWare
Jump to: navigation, search
(Autocreate 2013/10/03 Entry for Haynes_Lab:Notebook/David_Barclay_Undergrad_Training/Plasmid_Transformation)
 
(fix raw html notebook nav)
 
(5 intermediate revisions by one other user not shown)
Line 2: Line 2:
 
|-
 
|-
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes BioBrick Cloning</span>
 
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Haynes BioBrick Cloning</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"|
 
<!-- ##### 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. ##### -->
 +
=Continuation of Restriction Digests and Gel Electrophoresis=
 +
Restriction Digest Table<br>
 +
* Checked plasmid minipreps with EcoRI/PstI digests
  
 +
{| {{table}} border="1" cellspacing="3"
 +
<!-- Editing: the coding for this table is a bit more advanced. -->
 +
<!-- valign="top" aligns all the text in the first row to the top.  -->
 +
<!-- The | symbols on the next two lines start new cells in the same row. This does the same thing as ||, but you have to use | on new lines to set formatting for cells. -->
 +
<!-- bgcolor=#cfcfcf *colors* the *background* of the Reagent and Volume cells grey.  -->
 +
<!-- The next two "rowspan=7" cells span  all 7 rows in the table so that they can fit the "Expected" list and a gel image. rowspan is how you create merged rows in Wiki code. -->
 +
<!-- Replace Plasmid 1 and 2 with the names of your plasmids. Replace insert size and vector size with appropriate information for your plasmids. If you only have one Plasmid, delete all the text for Plasmid 2
 +
<!-- After you upload your gel image to OWW, replace "GelImage.jpg" with the name of your image file -->
 +
<!-- &nbsp; is ASCII code for an invisible space. -->
 +
|- valign="top"
 +
| bgcolor=#cfcfcf | Reagent
 +
| bgcolor=#cfcfcf | Volume
 +
| rowspan="7" | <u>Expected:</u><br>1. fshPCD = 186 bp, 4968 bp<br>
 +
| rowspan="7" | [[Image:Gel DKB 10-3-13.jpg|400px|Today's gel]]<br>15 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
 +
|-
 +
| DNA(plasmid) || 5.0 μL
 +
|-
 +
| 10X FD Buffer || 1.5
 +
|-
 +
| EcoRI || 1.0
 +
|-
 +
| PstI || 1.0
 +
|-
 +
| dH<sub>2</sub>O || 6.5
 +
|-
 +
| &nbsp; || 15 μL --> 37°C/ 15 min.
 +
|}
 +
'''Conclusion'''
 +
There were faint bands at the 186 position; however, they are harder to see on the picture. The 4968 bands for all 3 are very visible. Fairly successful digest.
 +
 +
'''DNA Sequencing Samples'''
 +
<!-- * signs create an automatically bulleted list. Replace 'date' with the date you submitted the samples -->
 +
<!-- The Order Number is shown at the top of your sequencing order form. Record in the indicated spot below -->
 +
* Submitted to DNASU on: 10/7/2013
 +
* Order Number: 8213
 +
<!-- # signs create an automatically numbered list. Replace Plasmid 1 & 2 with the names of your plasmids. Replace 'name' with the appropriate name of each primer. -->
 +
# Plasmid 1 (fshPCD) - Forward P001
 +
# Plasmid 1 (fshPCD) - Reverse P002
  
  

Latest revision as of 22:24, 26 September 2017

Owwnotebook icon.png Haynes BioBrick Cloning Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Continuation of Restriction Digests and Gel Electrophoresis

Restriction Digest Table

  • Checked plasmid minipreps with EcoRI/PstI digests
Reagent Volume Expected:
1. fshPCD = 186 bp, 4968 bp
Today's gel
15 μL/lane; 1% agarose; Ladder
DNA(plasmid) 5.0 μL
10X FD Buffer 1.5
EcoRI 1.0
PstI 1.0
dH2O 6.5
  15 μL --> 37°C/ 15 min.

Conclusion There were faint bands at the 186 position; however, they are harder to see on the picture. The 4968 bands for all 3 are very visible. Fairly successful digest.

DNA Sequencing Samples

  • Submitted to DNASU on: 10/7/2013
  • Order Number: 8213
  1. Plasmid 1 (fshPCD) - Forward P001
  2. Plasmid 1 (fshPCD) - Reverse P002