i have been trying to isolate DNA from mice blood from a long time using different kits. Presently i am using QIAGEN kit to isolate DNA . Initially i was getting nothing on the gel but after troubleshooting ( like decreasing dilution of blood, avoiding vortexing,less pipetting during gel load ) i am now getting some yeild and less shearing. However i want a clear sharp band and want to improve my yeild further.
Hence please suggest me possible ways. I have to get good amount of healthy DNA so that i can process it further for PCR .
Hi Ali,
Thank you for your advice.
Yes the human DNA isolation classical protocol might work but i think because the starting volumes for such protocol is 10 ml usually so this is not possible in case of mice also if i start with lower volumes then it would require further standardization and since i need to start early because of some urgency time is now a critical factor for me.
Also i tried with your other suggestion of working only with buffy coat ( rich in WBC) but till i get blood and start the protocol the spin down instead of giving me buffy coat gives serum even after adding heparin. ( time is critical i believe to get the buffy coat).
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