Haynes Lab:Notebook/Lab Traning - Pradyumna Kadambi/2013/06/28: Difference between revisions
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#Pellet was broken up using the vortex. | #Pellet was broken up using the vortex. | ||
#Added 100μL of lysis buffer to the centrifuge tube and inverted 4-6 times and waited 1 min. | #Added 100μL of lysis buffer to the centrifuge tube and inverted 4-6 times and waited 1 min. | ||
#Added 350μL of | #Added 350μL of neutralization buffer. Inverted the solution until it turns yellow, make sure no blue pockets are left and precipitate forms. | ||
# | #Centrifuged the solution at 16.1g for 2 mins. | ||
# | #Transfered the solution into a spin column and placed the spin column in a collection tube. | ||
# | #Centrifuged spin column/collection tube apparatus at 16.1g for 15 seconds. | ||
# | #The liquid in the collection tube was discarded. (Into the bleach container in the fume hood) | ||
#Spin column was put back into the collection tube, and 200μL of Endo Wash buffer was added to the spin column. | |||
#Centrifuged at 16.1g for 15 seconds. | |||
#400μL of Zippy Wash buffer was added to the spin column and the apparatus was centrifuged at 16.1g for 30 seconds. | |||
#Spin column was transferred to a new, clean 1.5 mL centrifuge tube. | |||
#30μL of the Zippy Elution Buffer was added to the column matrix (the white area), and waited 1 min. | |||
#Centrifuged at 16.1g for 15 seconds. (make sure to point the centrifuge tube caps away from the spin direction) | |||
#Discarded spin column, and labeled the centrifuge tube with initials, and plasmid.("PK KAH013") | |||
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Revision as of 19:48, 2 July 2013
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DH5α Incubation in Liquid Media 6/28/2013
DH5α Mini Prep with KAH013 6/28/2013
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