Difference between revisions of "Part:pSB6A1:Experience"

Revision as of 16:31, 1 October 2011

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of pSB6A1

User Reviews

UNIQf9648184eae39d85-partinfo-00000000-QINU

No review score entered. Username

This plasmid has been adapted to the version 5 of BioBrick vector backbones by the ETHZ 2011 iGEM team. This means, that the cassettes for the ORI and antibiotic resistance have been minimized (reduction of the size and transcriptional terminators flanking the prefix and the suffix have been added, to trancriptionally insulate the inserted parts from the vector backbone machinery and vice versa. This minimization yielded a reduction in size from 4022 bp to 2743 bp.

For comparison of the copy number of the plasmids, triplicates of OD normalized bacterial cultures containing the respective plasmids were miniprepped. The DNA concentration was then determined by an agarose DNA gel electrophoresis of EcoRI digests of the purified plasmids (Figure 1). This experiment showed a clear reduction in copy number of the minimized version of the original plasmid (Figure 2).

Figure 1: Agarose gel with 2-log-ladder loaded in in lane one, digests of triplicates of the EcoRI digests of pSB6A1 (lane 2-4) and pSB6A5 (lane5-7).

;

UNIQf9648184eae39d85-partinfo-00000002-QINU

@@ Line 17: / Line 17: @@
 This minimization yielded a reduction in size from 4022 bp to 2743 bp.
-For comparison of the copy number of the plasmids, triplicates of OD normalized bacterial cultures containing the respective plasmids were miniprepped. The DNA concentration was then determined by an agarose DNA gel electrophoresis of EcoRI digests of the purified plasmids.
+For comparison of the copy number of the plasmids, triplicates of OD normalized bacterial cultures containing the respective plasmids were miniprepped. The DNA concentration was then determined by an agarose DNA gel electrophoresis of EcoRI digests of the purified plasmids (Figure 1). This experiment showed a clear reduction in copy number of the minimized version of the original plasmid (Figure 2).
-This experiment showed a clear reduction of
 [[Image:Veeeeeeery final copy number test gel.png|300px|left|thumb|'''Figure 1: Agarose gel''' with 2-log-ladder loaded in in lane one, digests of triplicates of the EcoRI digests of pSB6A1 (lane 2-4) and pSB6A5 (lane5-7).]]