Difference between revisions of "IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/25"

From OpenWetWare
Jump to: navigation, search
(Entry title)
(fix raw html notebook nav)
 
(10 intermediate revisions by 3 users not shown)
Line 1: Line 1:
 
{|{{table}} width="800"
 
{|{{table}} width="800"
 
|-
 
|-
|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> iGEM Project name 1</span>
+
|style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]]<span style="font-size:22px;"> Week 8</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"|
 +
==Floor One==
 +
Streak purified all the isolated single Actinomycete colonies that appeared contaminated.
 +
 +
Many single colony plates were removed due to lack of growth.
 +
 +
All single colony plates were streaked to produce lawns (2 lawns per colony) in preparation for spore stocks.
 +
 +
After the conjugation plates were incubated at 30 °C overnight, they were overlayed with antibiotics - 1 ml of sterile water, 20 ul nalidixic acid and 100 ul kanamycin per plate. The plates were then incubated again overnight at 30 °C.
 +
 
==Floor Two==
 
==Floor Two==
The membrane was retrieved from the shaker and the blocking solution used was stored in the fridge. The membrane was washed with TBST and TBS solutions respectively for ten minutes. The membrane was incubated in Anti His HRP conjugate solution. After further washing the membrane was imaged using chemiluminescent detection. This gave no visible result.
+
The two ligation samples from the previous day were analysed by gel electrophoresis ''fig 2,'' alongside controls (2b2L and 3b1L) and the samples 2b and 2c from the 22nd July.
 +
[[Image:25 july.JPG|thumb|Fig 2: Analysis of Ligation reactions by gel electrophoresis. Lane 1 and 2 contain samples 1 and 2, lanes 3 and 4 controls, lane 5 and 6 samples from 22nd July (2b and 2c)repectively]]
 +
Transformation reactions were prepared using SOC media for the ligation reactions, J04450 plasmid and sample 3b from 11/07 (J04450 with Nde1 site).
 +
 
 +
The membrane was retrieved from the shaker and washed with TBST and TBS solutions respectively for ten minutes. It was then incubated in Anti His HRP conjugate solution. After further washing the membrane was imaged using chemiluminescent detection. This gave no visible result.
  
  

Latest revision as of 22:08, 26 September 2017

Igem-logo-150px.png Week 8 Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

Floor One

Streak purified all the isolated single Actinomycete colonies that appeared contaminated.

Many single colony plates were removed due to lack of growth.

All single colony plates were streaked to produce lawns (2 lawns per colony) in preparation for spore stocks.

After the conjugation plates were incubated at 30 °C overnight, they were overlayed with antibiotics - 1 ml of sterile water, 20 ul nalidixic acid and 100 ul kanamycin per plate. The plates were then incubated again overnight at 30 °C.

Floor Two

The two ligation samples from the previous day were analysed by gel electrophoresis fig 2, alongside controls (2b2L and 3b1L) and the samples 2b and 2c from the 22nd July.

Fig 2: Analysis of Ligation reactions by gel electrophoresis. Lane 1 and 2 contain samples 1 and 2, lanes 3 and 4 controls, lane 5 and 6 samples from 22nd July (2b and 2c)repectively

Transformation reactions were prepared using SOC media for the ligation reactions, J04450 plasmid and sample 3b from 11/07 (J04450 with Nde1 site).

The membrane was retrieved from the shaker and washed with TBST and TBS solutions respectively for ten minutes. It was then incubated in Anti His HRP conjugate solution. After further washing the membrane was imaged using chemiluminescent detection. This gave no visible result.