You can try cell lyses with CTAB follow by DNA precipitation with ethanol.
Normally it will give a yield of genomic DNA with ten-fold higher compare to the one using extraction kit.
By the way what kind of microalgae is that? Diatoms? Dinoflagellates? Cultured materials? Environmental samples?
If it's environmental samples, extraction with kits would be better.
Take algal suspension having at least 1gbiomass/litre. Use a bead beater to break up the cells. Quite difficult with tough cell wall filamentous green algae. After breaking, use the standard CTAB protocol to separate DNA. None of the available commercial kits give a quantitive extraction of DNA.
I have found the protocol used in this paper to work very well for every sample I have ever tried
from
Fawley and Fawley
J. Phycol. 40, 223–225 (2004)
r 2004 Phycological Society of America
DOI: 10.1046/j.1529-8817.2004.03081.x
DNA purification procedure. One to 2mL of algal
culture was centrifuged in a conical-bottom 2-mL
screw-top microcentrifuge tube at approximately
16,000 g for 1 min. The supernatant was discarded
and 200 uL extraction buffer (1M NaCl, 70mM Tris,
30mM Na2EDTA, pH8.6) added and vortexed
briefly. This suspension was then centrifuged at
16,000 g for 1 min, the supernatant discarded, and
200 uL fresh extraction buffer added. A quantity of
glass beads (G-8772, Sigma Chemical Co., St. Louis, MO, USA)
sufficient to fill the conical portion of the
centrifuge tube was then added, followed by 25uL
10% DTAB (Sigma Chemical Co.) and 200 mL chloroform.
A MiniBeadBeater was then used to disrupt the
cells, with agitation for 20 s at top speed. The mixture
was then centrifuged at 2000 g for 2 min to separate
the phases. If the cells were not well broken, as
evidenced by a cell layer at the interface of the
aqueous and organic phases and a lack of green
pigment in the organic phase, the MiniBeadBeater
step and centrifugation were repeated. The Mini-
BeadBeater step was never repeated more than once.
One hundred microliters of the aqueous phase was
removed to a 1.5-mL microcentrifuge tube, 500 mL
GeneClean salt (QBiogene, Carlsbad, CA, USA) was
added and mixed, and the resulting solution was
applied to a GeneClean Turbo (QBiogene) cartridge.
The cartridge was then used to purify the DNA
according the manufacturer’s instructions.
- Badges
- Science method