I am preparing C-TAB buffer and i read that addition of PVP (Polyvinylpyrrolidone) helps to remove phenolic compounds. Most of the times in protocols there is no additional information about average molecular weight of used PVP, but i found in some, where it was specified that it was PVP 40, 000, unfortunately in storage i have only PVP 360,000 can i use this or particles of polymer are too big ?
You will probably not find the molecular weight of PVP mentioned anywhere since it is a polymer. It will only mention the percentage or strength of the compound. If you want any particular concentration, you can go for dilution method for having strength of your choice.
Yes, it will effect in certain range of Mw. There are a big difference between 40 kDa and 360 kDa and unfortunately it will not depend on used concentration. You should find PVP near to 40 kDa.
Actually i have found in: "Protocols for Nucleic Acid Analysis by Nonradioactive Probes" ed Elena Hilario protocol for isolation from spores that use PVP 360:
Put 10 pretreated glass beads into an microcentrifuge tube containing the spore sample.
2. Add 0.2% β-mercaptoethanol and 1% PVP-360 to the CTAB buffer. Add 200 μL hot (65°C) CTAB buffer mixture to each tube (see Note 22)
3.Vortex for 1 min at high speed.
4. Incubate at 65°C for 30 min.
5. Add 200 μL chloroform/isoamyl alcohol (24:1).
6. Centrifuge at 14,000g for 15 min.
7. Precipitate with 2 volumes of 100% ethanol and leave sample at –20°C for at least 3 h. Repeat step 6.
8. Wash with 2 volumes of 70% ethanol, and repeat step 6. Discard supernatant.
9. Dry in a vacuum centrifuge.
10. Add 10 μL TE buffer. Store at 4 or –20°C.
Also I have found in Article Real-Time PCR for Quantification of Giardia and Cryptosporid...
that actualy PVP - 360 was more effective than PVP-40 in removing PCR inhibitors.
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