Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2013/01/07: Difference between revisions
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Caroline Hom (talk | contribs) (Autocreate 2013/01/07 Entry for Haynes_Lab:Notebook/Synthetic_Biology_and_Bioinformatics_for_Predictable_Control_of_Therapeutic_Gene2) |
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== | ==RNeasy Mini Kit== | ||
#Add 100μL of chloroform to each tube | |||
#Shake vigorously for 20 sec and let sit for 2 min | |||
#Centrifuge in cold room for 15 min @ 12,000G | |||
#Pipet off clear phase from organic phase into new tube (warning: do not get pink stuff in the tube!) | |||
#Add 70% ethanol into tube (1:1 ratio) | |||
#Transfer to collection tube (we have K562 1, K562 2, SK-N-SH 1, and SK-N-SH 2) | |||
#Spin tubes at top speed for 30 sec | |||
#Discard collected liquid and add 700μL of RW1 Buffer | |||
#Spin tubes at top speed for 30 sec | |||
#Put collection vial in a fresh collector tube and discard of the old one | |||
#Add 500μL of RBE Buffer | |||
#Spin tubes at top speed for 30 sec then discard of the liquid waste | |||
#Repeat the above two steps again | |||
#Discard of the waste and spin again | |||
#Transfer liquid to a fresh tube and add 30mL of RNase-Free H20 | |||
#Let tubes sit for 1 min | |||
#Spin tubes for 1 min | |||
#Store at -80°C | |||
Revision as of 11:47, 11 January 2013
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RNeasy Mini Kit
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