MagAttract 96 DNA Plant Core Kit (24)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Use of lyophilized leaf tissue	The amount of MagAttract bead suspension needed for each DNA sample can be reduced by 4 folds, from 20 μl (as required by the kit) down to only 5 μl

Upstream tips
Use of lyophilized leaf tissue
Protocol tips
The amount of MagAttract bead suspension needed for each DNA sample can be reduced by 4 folds, from 20 μl (as required by the kit) down to only 5 μl

Publication protocol

Protocol
1.Add one tungsten ball to the lyophilized leaf tissue (~10 to 40 mg) or dry seeds (10 seeds ~300 mg) in 2-ml microfuge tube. Place tubes in the grinding racks. Each rack holds 24 tubes. The two racks must hold equal number of tubes for balance.
2.Grind tissues at 28 strokes per second for 1 minute using TissueLyser II, repeat 3 more times, switching orientation each time for even grinding.
3.Add 750 μl extraction buffer to the ground tissue using multichannel pipette.
4.Cap the tubes and mix twice for 30 s at 20 strokes per second and incubate at 60°C for 1 hour.
5.Cool down tubes at room temperature for 5 min, then add 750 μl Chloroform:isoamylalcohol (24:1, v/v) to each tube, mix well, and centrifuge at 3000 g for 15 minutes.
6.Transfer aqueous layer (~500 μl) to a new set of labeled tubes.
7.Add 1 ml dilution buffer to the aqueous phase.
8.Mix well and incubate at 60°C for 30 minutes (Note: copious amount of precipitate of DNA-CTAB complex should be observed at the end of the incubation).
9.Centrifuge at 3000 g for 15 minutes and discard the supernatant.
10.Add 1 ml washing buffer to the pellet and soak it at RT for 30 minutes to remove access CTAB.
11.Centrifuge at 3000 g for 15 minutes and discard the supernatant.
12.Re-suspend DNA pellet in 100 μl high salt TE with RNase A and incubate at 60°C for 30 minutes.
13.Transfer the high salt TE DNA solution to 96-well micortiter plate, one sample per well.
14.Add 5 μl MagAttract suspension G solution to each well using a multichannel pipette.
15.Add 120 μl 100% ethanol to each well.
16.Cover the microtiter plate tightly with a silicone plate sealing mat and mix gently. Incubate it at room temperature for 5 minutes (to allow DNA adhere onto the surface of beads).
17.Place the DNA plate on Magnet B to hold the MagAttract beads and then pour off the ethanol solution.
18.Wash the beads three times with 200 μl washing buffer and air-dry the beads for 10 minutes at RT.
19.Add 100 μl TE to each sample well to re-suspend DNA.
20.Incubate at 60°C for 5 min and mix gently (to allow the bound DNA release into TE solution).
21.Place the plate on Magnet B and transfer DNA solution to a new 96-well plate.
22.Quantify DNA on a Nanodrop spectrophotometer or TECAN plate reader.

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Manufacturer protocol

Download the product protocol from Qiagen for MagAttract 96 DNA Plant Core Kit (24) below.

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