Aug 11, 2016
DIY Spin Column DNA Extraction
- Sarah Hessen-Schmidt1
- 1University of Southern California
Protocol Citation: Sarah Hessen-Schmidt 2016. DIY Spin Column DNA Extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.fjtbknn
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
Created: August 11, 2016
Last Modified: March 28, 2018
Protocol Integer ID: 3411
Keywords: diy spin column dna extraction, dna
Troubleshooting
Spin 500ul of bacteria broth culture at 10000g for 10 minutes. Pour off supernatant
Add 200ul Lysis buffer and 20ul Proteinase K. Vortex for 15 seconds and spin down
Incubate at 56 C for 1 hour
Add 200ul Binding Buffer and 200ul Ethanol to each sample. Vortex for 15 seconds and spin down
Transfer all 600ul of sample to a spin column and collection tube. Spin on high speed for 1 minute. Dispose of flow through
Add 500ul Wash Buffer. Spin on high speed for 1 minute. Dispose of flow through
Add another 500ul Wash Buffer. Spin on high speed for 1 minute.
Place the spin column in a new collection tube. Spin on high for 2 minutes
Place the spin column in a clean tube
Add 100ul TE buffer. Incubate at RT for 2 minutes.
Spin on high speed for 1 minute
Add another 100ul TE buffer. Incubate at RT for 1 minute. Spin on high speed for 1 minute.
Discard spin column. Extracted DNA is in tube.