Team:Cambridge/Protocols/Extraction from Filter Paper

Protocol Name

protocol description

• Cut out a portion of the plasmid-containing filter paper (half of the blot), and apply 50 μl elusion buffer (QIAGEN) in an eppendorf tube.
• Spin in the micro centrifuge for 1 min to elute the DNA from the paper.
• Add 250 μl of SOC to a 1.5 ml eppendorf tube of OD600 competent cells and transform with 1 μl of the plasmid DNA. When pipetting the plasmid, use the pipette tip to squeeze some fluid off the filter paper if necessary. At this point, the eluted DNA (with paper) was then stored in the freezer.
• Plate out 10 μl and 100 μl on LB agar plates

Miniprep extraction

• Transfer some bacteria from the colonies to liquid culture, and incubate for a further 16 hours.
• Perform a standard miniprep.

Theory

How it works

Practice

How to do it in the lab

{Standard layout for procedures is to use:

• <Procedure title - aka what you are doing>

1. <step 1>
2. <step 2>
   • <additional notes/important information regarding the previous step>

the text within the < > is what should be written, don't include < > in actual writeup :P

if in doubt see the gel electrophoresis protocol

}

Safety

The safety implication of the procedure.