Team:KAIST-Korea/Notebook/Diary/August
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== August 20 == | == August 20 == | ||
we went to KIB for cell lysis & DNA extraction. | we went to KIB for cell lysis & DNA extraction. | ||
- | # | + | # put 45mL of STAT1 solution into a cornical tube, put 45mL of FGFR solution into the other cornical tube. |
- | # | + | # add 6mL of SolutionI, then do vortexing. |
- | # | + | # add 6mL of SolutionII, then shake and roll. |
- | # | + | # add 6mL of SolutionIII, then shake and roll. |
- | # | + | # put each STAT1, FGFR solution in first column (10 min). |
- | # | + | # put 4mL of equilibrium buffer(QBT) into the second buffer. |
- | # | + | # put solution that passed by first column into second column. |
- | # | + | # add 20mL of washing buffer to the second column. |
- | # | + | # add 5mL of elution buffer to the second column. |
- | # | + | # add 5mL of isopropane in second column (5 min). |
- | # | + | # push the piston for DNA condensation |
- | # | + | # add TE buffer for elution |
- | # | + | # check DNA concentration using machine |
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After DNA extraction, we went to CMS builing for gel electrophoresis. | After DNA extraction, we went to CMS builing for gel electrophoresis. | ||
- | # | + | # prepare 1.2% of agarose gel, 0.5x TAE buffer |
- | # | + | # mix 1.5uL of STAT1, FGFR sample and 1uL of loading buffer |
# loading STAT1, FGFR, DNA marker (100V, (-)→(+)) | # loading STAT1, FGFR, DNA marker (100V, (-)→(+)) | ||
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[[Image:result0820_3.jpg|400px]] | [[Image:result0820_3.jpg|400px]] | ||
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- | We | + | We liked STAT1, FGFR DNA sample, and we will give these samples to bioneer next week. |
+ | <br> | ||
+ | == August 26 == | ||
+ | We went to Bioneer for communication. | ||
+ | <br> | ||
+ | Communicated topic | ||
+ | # Primer | ||
+ | #: We requested primer synthesis. But our list of requested primer was hard for them to recognize. Therefore they asked us to change and organize our request form. | ||
+ | # TA cloning | ||
+ | #: They asked us if we can do TA cloning in our lab. And we answered that we have no system that can manage TA cloning. Today, they offered us that they can do TA cloning for us. TA cloning is one additional way to store genes in safe way. | ||
+ | # Vector | ||
+ | #: We tell them that SAL1 is the only one that can be used in STAT1 cloning. But it is safe to have two enzyme sites. Therefore we need to check other enzyme sites such as BamH1, Sal1. | ||
+ | #pT218UHA12 vector | ||
+ | #: It is a way to change selection marker. Selection marker can be used first in this vector. Then after culture, we need to culture it in 5-FOA medium. Then selection marker can be deleted. If we use this method, we can use one selection marker twice, and this allow us to pick more vector in large choice. | ||
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Latest revision as of 05:22, 17 October 2010
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