We like to know the titers of our virus (I know - old school, but we do !) and with the advent of qPCR for titration, this has become an increasingly simple procedure. However, one of the time and money-consuming parts is the extraction of viral DNA. We use a Qiacube for this, but it is still resource-consuming.
As some of you may have seen, Oxford Expression Technology has recently launched a titration kit, where they lyse the baculovirus particles in a lysis buffer followed by a few heating steps in a thermocycler. Obviously, you need to buy the entire kit from them (£365.00 !) and the kit will contain far more than you need. All we need is the composition of the lysis buffer or the recipe for a buffer that will lyse baculovirus particles and release the DNA upon heating.
Here comes the question - does anybody know what is in the lysis buffer ? (I have asked OET just in case their contact was asleep at the wheel, but was not surprisingly told it was a secret, which probably means it is something very simple - but what ?).
Here is a link to the protocol: http://oetltd.com/client_media/medialibrary/2012/03/baculoQUANT_all-in-one_protocol_schematic.jpg
Thanks for any input,
NB: I have put this one on private - so whatever insights you may have are only available to group members
Thanks a lot, that was most useful - I'll have a careful read.
I also need Lysis buffer of baculovirus particles for titration and checking for insertion of the gene of interest.... so can anybody help me
can you please tell me where this document is uploaded to? I can't find the files section on this website.