Jun 14, 2022
DNA extraction (Zymo kit)
- 1KMU
Protocol Citation: Tsu-Chun Hung 2022. DNA extraction (Zymo kit). protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxz11ov8j/v1
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: In development
We are still developing and optimizing this protocol
Created: May 24, 2022
Last Modified: September 13, 2023
Protocol Integer ID: 63108
Abstract
DNA extraction (Zymo kit)
Sample Collection
Sample Collection
2m 30s
2m 30s
Add 200 µL of 1mm beads to 1.5ml enppendorf tube
30s
Add 200 µL of 0.5mm beads to 1.5ml enppendorf tube
30s
Add 600 µL of lysis solution to 1.5ml enppendorf tube
30s
Collect 9-10 µg of sample to 1.5ml enppendorf tube
1m
Sample crush
Sample crush
Put 1.5ml eppendorf tube in the mixmill for sample crush; at condition: 30 rpm/s, for 4mins 00:04:00
4m
Centrifugation
Centrifugation
Put 1.5ml eppendorf tube in centrifuge for centrifugation, at this condition:17.0 x g, 25°C, 00:03:00
3m
DNA purification
DNA purification
37m 30s
37m 30s
Add 600 µL of DNA binding buffer to 1st well of 96 deep well plate
30s
Add 25 µL of magnetic beads to 1st well of 96 deep well plate
Note
Shake the bottle and pipetting before using magnetic beads
30s
Add 200 µL of sample (lysate) from 15ml centrifuge microtube to 1st well of 96 deep well plate
Note
Usally add last
30s
Add 500 µL of Wash 1 to 2nd well of 96 deep well plate
30s
Add 500 µL of Wash 2 to 3rd well of 96 deep well plate
30s
Add 500 µL of Wash 2 to 4th well of 96 deep well plate
30s
Add 100 µL of water to 5th well of 96 deep well plate
30s
Put in the automated DNA extraction machine
34m