DNeasy from Quiagen work well for isolation of DNA from tissue: http://www.qiagen.com/Products/Catalog/Sample-Technologies/DNA-Sample-Technologies/Genomic-DNA/DNeasy-Blood-and-Tissue-Kit

Hi Sven:

So it's better to isolate during blastula stage or grastula stage?

Cheers,

Gan

Hello Gan,

I have no experience in that. I know that people isolate DNA with this kit also from mouse embryo material, but at which stage I don't know. I isolate only from adult material from human DNA with this kit.

Best

Sven

Hi Gan, I can recommend you the DNA Kit from MasterPure: http://www.epibio.com/products/nucleic-acid-purification-extraction-kits/dna-purification-genomic/masterpure-complete-dna-and-rna-purification-kit

It's easy to use it, a fast protocol and downstream applications (PCR, RT-PCR and Sequencing) are working very well. Cheers, Nadine

The gDNA purelink kit from Invitrogen works really well.

you can select one of the recommended kit mentioned as above. But,.you also can try to extract using conventional method as attached below.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3149498/pdf/1313.pdf

http://www.ncbi.nlm.nih.gov/pubmed/22907499

You click this link.

http://jhered.oxfordjournals.org/content/100/2/246.full

(From this article, genomic embryonic dna was extracted using Qiagen DNeasy kit)

Hopefully,it may help you to find the best way for extracting genomic dna from embryo.

Good Luck.

Hi, I have been using qiagen blood and tissue kit for blood samples. I have also used it for DNA extraction from bacteria. It works well

Two methods described here, for embryos we use the second method. Robust recovery of DNA from even just the head or tail of a 3 day old embryo.

Zebrafish DNA Lysis

From M. Sasaki (Kimmel Lab) Modified by G. Finch (Nicolson Lab)

For large amounts of tissue (e.g. tailfin clips) the faster "hotshot" method works great. For small amounts of tissue the traditional detergent/proteinase based method is more reliable.

FIN CLIPPING NOTE: If you are working with newly designed primers, it is recommended that you begin by clipping only a few (8?) fish to begin with. Designing primers and a PCR program that robustly produces a single band and gives a clean genotyping result can be tricky. Fin clipping takes time and changing the water (2X week) on fin clipped fish waiting for their genotyping results is time consuming, so make it easy on your self and totally optimize that PCR/sequncing before you clip a whole tank of fish.

Hotshot DNA Lysis

Solution A:

25 mM NaOH (50mg/50mL)

0.2 mM EDTA (2.9mg/50mL)

pH should be around 12

Solution B:

40mM Tris-HCl

pH should be around 5

Add tissue to 25uL of Solution A Incubate at 94oC for 20 min.

Add 25uL of Solution B Mix/Vortex

Use 2-5 uL for PCR, tissue may remain intact though DNA is present in solution.

Detergent/Proteinase DNA Lysis

Lysis Buffer 50ml:

500 ul 1M Tris pH 8.3

2500 ul 1M KCl

75 ul 1M MgCl2

750 ul 20% Tween-20

1500 ul 10% Nonidet-P40 (aka NP-40, Igepal) [10ml bottle of 10% Roche#11-332-473-001]

Add tissue to 25 ul of lysis buffer.

94oC - 20 minutes 4o -

Hold (Add 5ul 10mg/ml Proitenase-K)

55o - 3 hours

94o - 20 minutes

4o - Hold

Use 2-5 uL for PCR.