u want to isolate the human DNA from mosquitoes or DNA from mosquitoes. high molecular weight human genomic DNA isolated and DNA isolated from mosquitoes is qualitatively and quantitatively sufficient for DNA typing and could be helpful to identify individuals involved in certain cases of body violence or captivity. I think it may give some idea..

Use DNA extraction kits....there are many companies like qiagen, invitrogen....you can use DNA easy extraction kits....you have to cut the mosquitoes to very small parts ten fo;;ow the protocol

Yeh Mosaab Omar is absolutely give a correct protocol for DNA extraction kit....

Hi,

I suppose that you are collecting wild mosquitoes from the field, meaning that you should have a lot of sample to extract individually. Therefore I would not use Kits because it will cost you a lot of money.

1- Put each mosquito in 100microl of DNAZOl (Invitrogen) and grind them.

2-centrifuge 10min at 10,000g at 15°C

3-Transfer supernatant to new tube

4-add 50microliter of 100% EtOH Mix by inverting the tubes 5-8 times and hold 5 min at room temperature.

5- centrifuge 0min at 10,000g at 15°C and remove the supernatant.

6- Wash the DNA pellet with 1.0ml of 75% EtOH and invert tube 3-6 times to mix. Centrifuge 10min at 10,000g.

7-Repeat step 6.

8-Remove the remaining EtOH and dry pellet.

9-resuspend the dry pellet with 100microliter of free DNAse Water let it overnight at 4°C to maximize pellet rehydratation.

10- Storage at -80°C

As Mr. Mitri, the DNAzol is absolutely good method for a lot of sample. Additionally, I recommend you that if you use eukaryote universal primer, you should use small tissue like legs, not for whole body. Presumably in the whole body, there are many eukaryotes like human blood and many parasites.

Hi,

Use Wizard SV Genomic DNA Purification System kit (Promega).

Why from mosquitos

The following may be useful. Good Luck.

http://openwetware.org/wiki/Kafatos:genomic_DNA_extraction_from_individual_mosquitoes

http://www.mr4.org/Portals/3/Pdfs/Anopheles/3.12%2096%20well%20DNA%20extraction%20protocol%20v%201.pdf

http://journals.cambridge.org/action/displayAbstract;jsessionid=C8CC1C9FE6A515976E50C2AAB19AF07B.journals?fromPage=online&aid=763820

I agree with Chrstian good one try the same and let us know the results.personal thanks to Christy

You can also try this protocol. This works pretty fine with fruit fly. It should work fine with mosquitoes as well.

Lysis buffer Composition:

1M Tris-HCl pH 8.0……………100ul

0.5M EDTA……………………….20ul

3M NaCl………………………….330ul

10% SDS…………………………500ul

Sterile water upto…………..10ml

Procedure:

• Take single mosquito in an eppendorf tube and add 50ul lysis buffer.

• Homogenize in Ice

• Add equal volume of Phenol

• Centrifuge at 8000rpm for 10 mins

• Take the supernatant

• Add equal volume of Chloroform-Isoamylalchohol(24:1) mixture

• Centrifuge at 8000 rpm for 10 mins and take the supernatant

• Add 1/10th volume of 3M sodium acetate

• Add 2.5 volume of chilled Ethanol

• Keep at -20°c overnight

• Centrifuge at 10,000rpm for 5 mins

• Decant the supernatant

• Wash with 70% alchohol

• Centrifuge at 10,000rpm for 5min

• Decant the supernatant

• Air dry the pellet

• Dissolve the pellet in 50ul of sterile water.

Good luck.

Thank u very much .....!!!!!

we usually used Qiagen DNeasy Blood & Tissue Kit (Cat. No:69504) to extract the genomic DNA from mosquitoes.

Thanks all dears..............

use Qiagen DNeasy Blood & Tissue Kit

gd luck....

Number of companies supply DNA extraction kits[qiagen, invitrogen]

http://openwetware.org wiki/Kafatos:genomic_DNA_extraction_from_individual_mosquitoes lalitha kabilan