Team:Newcastle/11 August 2010

From 2010.igem.org

Revision as of 12:42, 12 August 2010 by Shethharsh08 (Talk | contribs)

iGEM Homepage Newcastle University BacillaFilla Homepage Image Map

Contents

Gibson cloning of the rocF BioBrick

Aim

The aim of this experiment is to assemble the fragments we extracted yesterday in order to construct the rocF BioBrick and clone it into pSB1C3 in a single step isothermal reaction.

Materials and Protocol

Please refer to: Gibson cloning for materials required and the protocol for the reaction.

Result

Subtilin Immunity BioBrick

Aims

Our aims for today are to run the gel electrophoresis to check whether we have the correct fragement sizes on the four parts that are we amplified yesterday. If the PCR worked, we will then perform gel extraction and then perform another gel electrophoresis for the extracted gel in order to obtain our BioBrick parts.

Newcastle Loaded Gel for Subtilin Immunity BioBrick.JPG

Materials and protocol

Please refer to the gel electrophoresis and the gel extraction protocols.

Results

Newcastle Subtilin Immunity Gel 1.jpg

Figure #: Gel electrophoresis of the PCR products of the parts required for the subtilin immunity BioBrick.

  • Lane 1: 1 kb DNA ladder
  • Lane 2: Plasmid Vector (pSB1C3)
  • Lane 3: Promoter and RBS (pVeg-SpoVG)
  • Lane 4: spaIFEG Gene Cluster
  • Lane 5: Double terminator
  • Lane 6: 100 bp DNA ladder

Discussion

Plasmid Vector (lane 2), Promoter & RBS (lane 3) and Double terminator (lane 5) showed up. spaIFEG PCR tube (lane 4) did not show up. We think that this occurrence was due to the Tm error (it should be 63°C as opposed to 46°C). Therefore, another PCR and gel electrophoresis were performed.

Transformation of hyperspank and spoVG

Aim

To transform competent E. coli DH5α with hyperspank and spoVG.

Materials and Protocol

Please refer to transformation of E. coli.


Go back to our main Lab book page

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