Hey there,
I've been trying to purify these influenza samples, however, I've come across 2 problems. I've repeated the procedures twice and have come across the same issues both times (first time thinking some error had occurred).
Sample 1: After PCR, product is visualised on gel to see if there is a correct product size. There is but also some non specifics, so I cut the band out. After gel purification, the band is now larger bp than original. Size of PCR product is 916bp. However, after purification, it is now well above 1000 bp.
Sample 2: After gel purification, the product has disappeared.
I know it is not an error of the kits (i.e forgetting to add ethanol to wash buffer or forgetting to incubate) as I have used these same kits to purify many samples before it. So these are perhaps sample-specific problems.
I have run these samples using both the Qiagen Gel Purification kit and the Intron Purification kit. Both times have encountered the same problem.
Any recommendations on what I should try?
Thank you so much!
If that might help, you could try a purification of the PCR product itself first and then run a gel to see if you get something different.. One possibility for the different gel results for samples 1 and 2 after gel purification is the difference in "intensity" of non-specific bands before the gel purification. Hope that might help. Best of luck! Alina
If you use the Qiagen Gel Purification kit it really helps when you warm the Elution buffer up to 37 degree. But this just help to increase the yield of eluted DNA.
For the first question did you use the gel with the same percentage of agarose and the same marker from the same company?
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