Team:Newcastle/22 June 2010

From 2010.igem.org

(Difference between revisions)
(Transformation protocol)
(Nanodrop Protocol)
 
Line 100: Line 100:
{|
{|
|-
|-
-
|[[Image:Newcastle_nanodrop_1.jpeg|thumb]]
+
|[[Image:Newcastle_nanodrop_1.jpeg|thumb|A typical curve to be achieved from a nanodrop spectrophotmeter]]
-
|[[Image:Newcastle_nanodrop_2.jpeg|thumb]]
+
|[[Image:Newcastle_nanodrop_2.jpeg|thumb|Phil analyzing a sample on the nanodrop machine]]
-
|[[Image:Newcastle_nanodrop_3.jpeg|thumb]]
+
|[[Image:Newcastle_nanodrop_3.jpeg|thumb|Analyzed data on the screen]]
|-
|-
|}
|}

Latest revision as of 20:42, 21 October 2010

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Contents

Aims

The aim of today's Lab practice was to extract the plasmids for GFP and RFP, digest the 2 plasmids and extract the 2 inserts and 1 of the backbones (vector)from an agarose gel. From this a ligation was set up.

Materials and Protocol

QIagen Miniprep Kit Using a Microcentrifuge for Plasmid Extraction

Please refer to: Qiagen Minipreps for materials required and protocol.

Digest

Please refer to: Restriction digests for materials required and protocol.

Gel extraction

Please refer to: Gel extraction for materials required and protocol.

Agarose Gel Electrophoresis

Please refer to: Gel electrophoresis for materials required and protocol.

Dr Wendy demostrating how to use gel electrophoresis machine
Gel tank
Deena pouring hot molten gel

Cut gel out

Putting the gel on the geldoc
Preparing to cut the gel
Gel illuminated by the UV light which is emitted by the geldoc

We cut the inserts which were about 900bp and we use the backbone from the plasmid consisting rfp.

QIAquick Gel extraction kit

Phil cutting the gel
Transferring the cut gel part into a 2 ml microfuge tube

Set up ligation

Reagents 1:3(μl) 1:5(μl) Vector(μl)
V 0.8 0.8 1
G 2.7 4
R 5.4 7.7
LT4 1 1 1
LB 1.1 1.5 1
H2O 0 0 7
Total Volume 11.0 15.0 10.0

Transformation protocol

Please refer to: Transformation of E. coli for materials required and protocol.

Nanodrop Protocol

Please refer to: Nanodrop Spectrophotometer for materials required and protocol.


A typical curve to be achieved from a nanodrop spectrophotmeter
Phil analyzing a sample on the nanodrop machine
Analyzed data on the screen

Outcome

Home Team Official Team Profile Project Parts Submitted to the Registry Modelling Notebook Safety
Newcastle University logo.png    Newcastle cbcb logo.pngNewcastle Biomedicine logo.gif    Team Newcastle CEG logo.gif
Newcastle iww logo.jpg  UNIPV Pavia Logo.gif  Newcastle BBSRC.gif    Newcastle Genevision logo.png Newcastle WelcomeTrust.jpg
FaceBook Icon