Latest revision as of 23:41, 27 October 2010

Home

Team

Problem

Gel electrophoresis of subtilin immunity part fragments

Aims

The aim of this experiment is to do gel extraction for the spaIFEG gene cluster amplifed on the 12th August, 2010 and the Plasmid Vector, Promoter & RBS and Double terminator amplifed on the 11th August, 2010. Finally, NanoDrop will be performed for all the fragments.

Materials and protocol

Please refer to the gel extraction and NanoDrop protocols. Instead of using the centrifuge to bind the DNA from our sample to the QIAquick column, a vacuum manifold was used instead. It is the first time it had been used. The advantage of this was that the extraction could be carried out all at the same time.

Results

Vector	Promoter	Immunity gene cluster	Double terminator
21.3 µl/ml	28.8 µl/ml	27.0 µl/ml	27.5 µl/ml

Table 1: Nanodrop spectrophotometer results. Table represents the amount of plasmid present in µl/ml quantity.

Discussion

The concentration of the fragments range from 21.3 µl/ml to 28.8 µl/ml. This is acceptable as during the gel extraction step, some of the DNA would be lost.

Conclusion

We have successfully extracted all the parts required for the Subtilin Immunity BioBrick. The Gibson cloning method will be used next week.

Go back to our main Lab book page

@@ Line 5: / Line 5: @@
-=Gel electrophoresis of subtilin immunity BioBrick fragments=
+=Gel electrophoresis of subtilin immunity part fragments=
 ==Aims==
@@ Line 21: / Line 21: @@
 ==Results==
-The results from the Nanodrop Spectrophotometer of the four parts are as follows:
-*'''Vector''' - 21.3 ng/μl
-*'''Promoter''' - 28.8 ng/μl
-*'''Coding sequence''' - 27.0 ng/μl
-*'''Terminator''' - 27.5 ng/μl
 {|border=1
@@ Line 44: / Line 38: @@
 ==Discussion==
-We chose the three ''spaIFEG'' gene (part 3) PCR tubes, using Tms 51°C, 56°C and 61°C because they were the mid-ranged Tms. Gel extraction was then performed, by firstly cutting the gels under U.V light, and then apply the protocol for completion.
+The concentration of the fragments range from 21.3 µl/ml to 28.8 µl/ml. This is acceptable as during the gel extraction step, some of the DNA would be lost.
 ==Conclusion==
-The results from the NanoDrop reading suggests that our results were positive, which indicates that we have successfully extracted all the parts required for the Subtilin Immunity BioBrick. Gibson cloning method will be used next week.
+We have successfully extracted all the parts required for the Subtilin Immunity BioBrick. The Gibson cloning method will be used next week.

Team:Newcastle/13 August 2010

From 2010.igem.org