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BIOTEC Dresden/Notepad/26 August 2010
From 2010.igem.org
PCR amplification of backbone was done with the primers ASP and SP and with a slightly varied PCR program this time. The PCR samples were purified and then run on an agarose gel. In addition to the bands corresponding to the right molecular weight, there were less intense bands corresponding to molecular sizes not relating to the backbone size. This maybe due to some unspecific binding which has to be looked into.
Restriction digest for the following parts was done for 1 hour and 15 minutes.
4a, 9b, 6e, 14f, 20f, 21f
The digests were purified and run on an agarose gel. The bands were totally as per the expected molecular weight.
Fusion Protein
The PCR reactions for aiiA and the 10aa linker were set up. All reactions were purified using the Quiagen PCR Purification Kit. All concentrations ranged from 150 - 580 ng/ul. The short linker fragments were precipitated using Ethanol precipitation.
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