Haynes Lab:Notebook/Lab Traning - Pradyumna Kadambi/2013/06/28: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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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") | |||
==Spectrophotometer analysis== | |||
#Turn on spectrophotometer. | |||
#Open the program "Gen 5 2.0" (make sure spectrophotometer is on FIRST). | |||
#Click "Take 3 Plate Application," and select the "Nucleic Acid Quantification" routine. | |||
#Clean loading plate with Kimtech wipes and DI water. | |||
#First, load a 2μL blank of Zippy Elution buffer, then load the sample. (Plate is magnetic, so remember to hold it down when closing the loading plate) | |||
#Mark the blank and the sample properly on the computer, and click "approve." | |||
#After gathering data, clean loading plate with Kimtech wipes and DI water. | |||
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Latest revision as of 22:53, 26 September 2017
Project name | Main project page Previous entry Next entry |
DH5α Incubation in Liquid Media 6/28/2013
DH5α Mini Prep with KAH013 6/28/2013
Spectrophotometer analysis
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