Team:Brown/Notebook/Protocols
From 2008.igem.org
(Difference between revisions)
NeilParikh (Talk | contribs) (→Gel Extraction) |
NeilParikh (Talk | contribs) (→Calf CIP - Calf Intestinal Alkaline Phosphatase) |
||
Line 93: | Line 93: | ||
#Elute gel extract in 30 microliters 10 mM Tris-HCl Buffer (Buffer EB) | #Elute gel extract in 30 microliters 10 mM Tris-HCl Buffer (Buffer EB) | ||
#CIAP 10x Reaction Buffer and CIAP enzyme should thaw on ice until used. Add 4 microliters CIAP 10x Reaction Buffer and 1 microliter CIAP enzyme to the 34 microliters from the gel extraction. The total volume should be 39 microliters. | #CIAP 10x Reaction Buffer and CIAP enzyme should thaw on ice until used. Add 4 microliters CIAP 10x Reaction Buffer and 1 microliter CIAP enzyme to the 34 microliters from the gel extraction. The total volume should be 39 microliters. | ||
- | # | + | #Incubate in 37 deg C for 30 minutes. |
- | # | + | #Add 1 microliter CIAP enzyme. Incubate in 37 deg C for 30 minutes. |
*An Ethanol Precipitation is performed on the vector after the Calf CIP procedure and on the insert after Gel Extraction. The Ethanol Precipitate is usually placed in -80 deg C overnight OR for 1 hour coupled with a 30 minute centrifuge at 4 deg C. | *An Ethanol Precipitation is performed on the vector after the Calf CIP procedure and on the insert after Gel Extraction. The Ethanol Precipitate is usually placed in -80 deg C overnight OR for 1 hour coupled with a 30 minute centrifuge at 4 deg C. |
Latest revision as of 21:31, 29 October 2008
ProtocolsMiniprep Prodedure
Restriction Digest
Gel Electrophoresis
Ligation Reaction Procedure
Transformation Procedure
Ethanol Precipitation
Gel Extraction
The contents of the tube should be yellow after incubation.
Calf CIP - Calf Intestinal Alkaline Phosphatase
|