Home › Genetics / Genomics › DNeasy visual protocol
Steps
  1. 1 Prepare buffers and check for precipitates 00:39
  2. 2 Lyse sample with proteinase K and buffer 01:06
  3. 3 Add ethanol and load onto spin column 01:38
  4. 4 Perform first wash with buffer aw1 02:19
  5. 5 Perform second wash with buffer aw2 02:40
  6. 6 Elute DNA with buffer AE 03:22
  7. 7 Optional re-elution for higher DNA yield 03:45
  8. 8 Verify DNA purity and downstream applications 04:08
Genetics / Genomics QIAGEN

DNeasy visual protocol

Protocol
Difficulty
intermediate

Steps

1
Prepare buffers and check for precipitates

Add the recommended amount of ethanol to buffer aw1 and aw2 concentrate bottles before first use. Check buffer AL and ATL bottles for any precipitates and redissolve if present.

▶ 00:39
2
Lyse sample with proteinase K and buffer

Add proteinase K to the sample, then add buffer AL for blood samples or buffer ATL for tissue samples and vortex thoroughly. For blood samples, incubate at 56°C for 10 minutes.

▶ 01:06
3
Add ethanol and load onto spin column

Add 200 microliter of ethanol and vortex thoroughly. Pipet the entire mixture into a DNeasy spin column and centrifuge for 1 minute, then discard the flow-through and collection tube.

▶ 01:38
4
Perform first wash with buffer aw1

Place the spin column into a new collection tube, add 500 microliter of buffer aw1, and centrifuge for 1 minute. Discard the flow-through and collection tube.

▶ 02:19
5
Perform second wash with buffer aw2

Add 500 microliter of buffer aw2 to the column and centrifuge for 3 minutes at 13,000 RPM. Discard the flow-through and collection tube, then centrifuge for an additional minute to remove residual ethanol and salts.

▶ 02:40
6
Elute DNA with buffer AE

Place the spin column into a new microcentrifuge tube, add 200 microliter of buffer AE, and incubate for 1 minute at room temperature. Centrifuge for 1 minute to extract the purified DNA.

▶ 03:22
7
Optional re-elution for higher DNA yield

For increased DNA yield, repeat the elution step using the purified DNA already collected. This increases the elution efficiency for small volumes.

▶ 03:45
8
Verify DNA purity and downstream applications

The purified DNA is free from PCR inhibitors and ready for use in standard, multiplex, and real-time PCR as well as sequencing applications.

▶ 04:08
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