Hi all, i would like to know the what is the minimum amount used to lyophilized the LAMP reagents. My lab is using Telstar freeze-dryer and she mention that usually they used 5mL and they are food sample products. In my case, the reagents i will lyophilize include deglycerol Bst DNA polymerase, fluoroscent dye, primer, dNTPs and stabilizer and overall the total amount is just 15ul (acc to the protocol i read through). Is it possible to lyophilized less than 1mL or maybe roughly 100ul of the LAMP reagents?
Sarah - sure, it's possible, folks do it all the time.
You have to have a freeze dryer that actively freezes the sample on a shelf before applying the vacuum, and then keep the sample frozen with the shelf. If you have a basic lab model (flasks and a tree) I think that this will not work well even if you pre freeze the sample.
Things to watch out for:
If you use plastic ware to contain the sample, sometimes injection moulded plastics have things in the plastic or chemicals used in the production process (waxes or silicones to help the plastic part com off the mould) that will affect the drying process
pH shift when freezing a buffer salt, phosphate buffers can be especially prone to this.
Kind regards
Rob
Rob Darrington thanks for your reply. i come across a video which they lyophilize inside a small tube https://www.youtube.com/watch?v=-INsuz3H1M0
and am wondering is it possible to do in such a way? Im actually planning to lyophilize inside a PCR tube consisting of 15ul for one reaction but i was told that it will end up nothing after lyophilize; so instead of preparing 1 reaction, i prepare 10 reactions which give a total amount of 150ul. Any more suggestion u can give for this?
Many companies producing test kits do lyophilise inside PCR tubes and plates. The equipment is a production system with freezing shelves to control the temperature of the sample throughtout the process, also, the check very carefully the formulation that is used for freeze drying. It is possible, with care, and the correct equipment.
Kind regards,
Rob
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