10 September 2013 1 1K Report

Does anyone have SDS based DNA isolation protocol from Aedes aegypti mosquito larvae or larvae of any related organisms? I have to isolate DNA from freshly emerged mosquito larvae and older. I have tried this protocol once:

1: grounded the larvae in TE pH 8.0

2. Added 10 uL of 10mg/mL RNase A. Incubated at 37'C for 30 min

3. Added 25 uL of 10mg/mL Proteinase K. Incubated at 55'C for 15 min.

4. Added 30 uL of 20% SDS. Incubated at 60'C for 30 min.

5. Continued with chloroform:isoamyl alcohol (24V:1V) extraction.

6. Resuspended in 30uL of TE buffer.

When I used picodrop to quantify the amount of DNA, I had low yield. Moreover, the absorption was funny looking. I had large flat peak ranging from about 200-500 nm. Does anyone have guess on what might have happened? I appreciate the help in advance.

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