Can anyone tell me what I should use to dissolve TPCK powder to be used in DMEM to culture Influenza? I`ve been using 100nM tris-HCL and just wanted to che ck this is correct
Cheers
Thanks Hershey
Do you know what the difference is between tpck trypsin and
Tpck-treated trypsin. Some papers state they used the former and some the latter, I presume there is no difference
Cheers
Hershna
What time post infection would you passage influenza virus and what volume would you put onto a confluent MDCK cells in a T75 fas?
A lot of your questions are answered in the WHO publication http://www.who.int/influenza/gisrs_laboratory/manual_diagnosis_surveillance_influenza/en/
You can e-mail me directly for additional help. [email protected].
Thomas
Mark, I usually just make mine directly in DMEM and filter. Then I store @-70C in small aliquots. Pull out an aliquot and make fresh in your medium.
You can dilute the TPCK-treated trypsin in DMEM directly, then filter it (0.22 μM), aliquot it in 500 μl (or any other amount that is convenient for use) and store it at -20°C or below for maximum 1 year.
Only thaw the aliquot that you will need, do not thaw-freeze back and do not prepare your "virus growth medium" with the added TPCK-trypsin too long before use because the trypsin is quite sensitive.
Thanks everyone, this is almost like a worlwide effort to get my project off the ground!
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