I am trying to prepare multiple samples with Invitrogen's Sequalprep long range PCR kit for sequencing on a 454. Does anyone have any experience of this, for example how much do you need to quality control your PCR products (for specificity etc) before quantification and pooling?
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Bad efficiency
We've tried using this kit for enrichment of regions around 10-12kb. The efficiency for this was near zero. We downscaled it down to 3-7kb and are starting to get amplification efficiency around 30-50%.
For our purposes this kit is waste of money, especially because of i) the price, and ii) nonexistent customer support. We wrote several times to speak to people who actually KNOW how this kit works, but we got general answers about how to design primers.....
Well, the best option for us would be raindance, but who can afford this...
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