840:153g:Projects/project24/2012/10/11: Difference between revisions
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This week on Tuesday, we did Ag-arose gel electrophoresis for our controls and two DNA samples. This time we found bands in our Positive control which was satisfactory result because we didn't found bands of Positive control in our previous experiment which we performed last week. However, we did found thin bands on our negative control but we assume that those bands were of primers oligos. So, we decided to do PCR for our DNA samples on Thursday. | This week on Tuesday, we did Ag-arose gel electrophoresis for our controls and two DNA samples. This time we found bands in our Positive control which was satisfactory result because we didn't found bands of Positive control in our previous experiment which we performed last week http://openwetware.org/wiki/Image:PIC_4_-_Copy.jpg. However, we did found thin bands on our negative control but we assume that those bands were of primers oligos. So, we decided to do PCR for our DNA samples on Thursday. | ||
On Thursday, we performed PCR for our both DNA samples( DNA1 and DNA 2) with 6 different temperature ranges(44.8ºC, 49.3ºC,55.2ºC,59.1, 63 and 65ºC) and controls in lowest temperature i.e, 40ºC.This time we thawed both the primers and Plasmid DNA completely which we didn't on previous PCR reaction. We also used only non-extended primers whereas, last time we used both( extended and non-extended). We also increased the volume of our DNA sample by 3 times i.e we used 3ul this time.We run our samples in to thermal cycler for over night and we will take those out of the cycler tomorrow, 10/12 morning and store it on -20ºC. | On Thursday, we performed PCR for our both DNA samples( DNA1 and DNA 2) with 6 different temperature ranges(44.8ºC, 49.3ºC,55.2ºC,59.1, 63 and 65ºC) and controls in lowest temperature i.e, 40ºC.This time we thawed both the primers and Plasmid DNA completely which we didn't on previous PCR reaction. We also used only non-extended primers whereas, last time we used both( extended and non-extended). We also increased the volume of our DNA sample by 3 times i.e we used 3ul this time.We run our samples in to thermal cycler for over night and we will take those out of the cycler tomorrow, 10/12 morning and store it on -20ºC. | ||
Unfortunately, we are out of our DNA 1 sample, so we decided to cut the gel and collect DNA 1 if it showed good band in the Agarose gel which we will be performing on our next lab. | Unfortunately, we are out of our DNA 1 sample, so we decided to cut the gel and collect DNA 1 if it showed good band in the Agarose gel which we will be performing on our next lab. | ||
Revision as of 13:59, 29 November 2012
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This week on Tuesday, we did Ag-arose gel electrophoresis for our controls and two DNA samples. This time we found bands in our Positive control which was satisfactory result because we didn't found bands of Positive control in our previous experiment which we performed last week http://openwetware.org/wiki/Image:PIC_4_-_Copy.jpg. However, we did found thin bands on our negative control but we assume that those bands were of primers oligos. So, we decided to do PCR for our DNA samples on Thursday.
On Thursday, we performed PCR for our both DNA samples( DNA1 and DNA 2) with 6 different temperature ranges(44.8ºC, 49.3ºC,55.2ºC,59.1, 63 and 65ºC) and controls in lowest temperature i.e, 40ºC.This time we thawed both the primers and Plasmid DNA completely which we didn't on previous PCR reaction. We also used only non-extended primers whereas, last time we used both( extended and non-extended). We also increased the volume of our DNA sample by 3 times i.e we used 3ul this time.We run our samples in to thermal cycler for over night and we will take those out of the cycler tomorrow, 10/12 morning and store it on -20ºC.
Unfortunately, we are out of our DNA 1 sample, so we decided to cut the gel and collect DNA 1 if it showed good band in the Agarose gel which we will be performing on our next lab.
