Hello everyone,
I'm using ELISA for plant viruses first time and following DSMZ protocol. I need to prepare some buffers called
1. PBS-Tween:PBS+0.5 ml Tween 20/liter (this much only given in the protocol). I need to know how much PBS to be used in 1 L and what is that 1L is H2O or the PBS itself.
2. Sample extraction buffer: PBST+2% PVP
How much of PBST and 2% PVP in what quantity.
3.Conjugate buffer: PBST+2%PVP+0.2%egg albumin
how much of PBST, 2% PVP and 0.2% egg albumin in what quantity.
Can anyone help me out to prepare above buffers.
Thanks Chandrika
Dear Chandrika,
It's very simple.
I prepare 0.5 L PBST.
1. you need to dissolve 0.25 ml Tween 20 at 0.5 L PBS, which has been prepared in a specific molarity.
2. PBST + 2% PVP+: 10 g of PVP is dissolved in 400 ml of PBST and
after dissolving the add the remaining amount of PBST to 0.5 L.
3. Conjugate buffer:0,2 g of egg albumin is dissolved in 100 ml of PBST + 2% PVP (prepared above).
Dear Chandrika M N
Pls. find the attached files you may need in your virology lab.
page 65 in: S. E. Albrechtsen. 2006.Testing Methods for Seed-transmitted Viruses: Principles and Protocols. CABI, Pp. 268.ISBN 1845931483, 9781845931483
Hoping this will be helpful
Regards
Prof. Houda Kawas
Also
Page 1148 in: Ronald M. Atlas. 2010. Handbook of Microbiological Media, Fourth Edition CRC Press, Pp.2040 ISBN1439804087, 9781439804087
https://www.dsmz.de/catalogues/catalogue-plant-viruses-and-antisera/faq.html
https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium1132.pdf
https://www.dsmz.de/uploads/media/DAS-ELISA-Pepino.pdf
https://www.dsmz.de/uploads/media/TAS_ELISA_01.pdf
http://www.cabri.org/guidelines/plant_virus/pcv204ap2.html
http://www.protocol-online.org/biology-forums/posts/34677.html
Thanks a lot @ Kashimova and Kawas, this has helped me a lot for my calculation.
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