Team:NRP-UEA-Norwich/Week5

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. Rachel and Russell calculated the expected length of plasmid and found that the plasmids on the agarose gel were completely different to expected. Decided to carry out a restriction digest of the plasmids to remove the inserts and then run those on a gel to find out if they are as expected. After a nanodrop we found that there was a very small amount of DNA and likely too little to work with; mini-prepped the second samples for each biobrick and nanodropped them.
. Rachel and Russell calculated the expected length of plasmid and found that the plasmids on the agarose gel were completely different to expected. Decided to carry out a restriction digest of the plasmids to remove the inserts and then run those on a gel to find out if they are as expected. After a nanodrop we found that there was a very small amount of DNA and likely too little to work with; mini-prepped the second samples for each biobrick and nanodropped them.
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. Rachel transformed new E. coli with the original biobricks in order to produce a new stock of DNA as we had left gaps between using the bacteria and DNA previously which could account for the very low levels of DNA we had experienced in the nanodrops.
==Day 3==
==Day 3==

Revision as of 10:33, 8 August 2012

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NRP UEA iGEM 2012

 

Welcome to the NRP UEA iGEM 2012 Wiki Lab Book

Please choose the relevant link to access our diary of that week!

Week Zero | Week One | Week Two | Week Three | Week Four | Week Five | Week Six | Week Seven | Week Eight | Week Nine | Week Ten | Week Eleven | Lab Protocols | Experiments

Contents

Week 5

Day 1

. Rachel and Russell ran their RFP/eCFP/AraC isolated plasmids on an agarose gel to see if they were there. The results of the gel seemed unusual.

Day 2

. Rachel and Russell calculated the expected length of plasmid and found that the plasmids on the agarose gel were completely different to expected. Decided to carry out a restriction digest of the plasmids to remove the inserts and then run those on a gel to find out if they are as expected. After a nanodrop we found that there was a very small amount of DNA and likely too little to work with; mini-prepped the second samples for each biobrick and nanodropped them.

. Rachel transformed new E. coli with the original biobricks in order to produce a new stock of DNA as we had left gaps between using the bacteria and DNA previously which could account for the very low levels of DNA we had experienced in the nanodrops.

Day 3

. Rachel checked the transformed bacteria and found that some had grown but others hadn't grown quite as much; left for a few more hours to grow.

. Russell carried out a restriction digest of each isolated plasmid hopefully containing the RFP, eCFP or AraC biobricks. He did one digest of each plasmid with just EcoR1 to linerarise the plasmid, and one digest of each plasmid with EcoR1 + Pst1 to cut out the insert. These digested plasmids will then be run on an agarose gel in order to validate the DNA that are in the biobricks due to the unusual results previously; if they are fine then the biobricks transformed into E. coli by Rachel will be used to produce new biobricks and carry out quantitative experiments.

Day 4

Day 5

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