chip assay dna pptn - (Oct/10/2007 )
Hello all,
I did a Chip this week and in the final step where we are supposed to precipitate the DNA using 100% Ethanol, by mistake I used 70%Ethanol....I realized my mistake but coudlnt do much about it then and added ~300-400ul of 100% ethanol and 2ul of glycogen and put my samples at -20C o/n...Im wondering if I will get good DNA prepitation...is there something else i can try to aid the DNA pptn..pls help!!
I think you should be fine if you have compensated the concentration of ethanol by adding additional 100% ethanol. For DNA precipitation, usually you add 2x 100% ethanol to get a final concentration of 67%. So you can more 100% ethanol to make the same conc. also you need to add more salt to match the new volume.
Actually chip DNA can be purified using qiagen PCR purification kit which is much easier.
I did a Chip this week and in the final step where we are supposed to precipitate the DNA using 100% Ethanol, by mistake I used 70%Ethanol....I realized my mistake but coudlnt do much about it then and added ~300-400ul of 100% ethanol and 2ul of glycogen and put my samples at -20C o/n...Im wondering if I will get good DNA prepitation...is there something else i can try to aid the DNA pptn..pls help!!
I did add 100% EtOH but could add only about 400ul since im using 1.5 ml eppendorfs-
I was thinking of transferring them to 2ml eppendorf tubes- adding another 500-600 ul 100% Etoh-but in that case since its better to centrifuge in 1.5ml eppendorf tubes- can i centrifuge them twice tomorrow- centrifuge 1ml first, discard the sup and then add another 1ml to that tube to cent again- will that be ok?
why not split it into two 1.5 ml tubes?