In common ELISA for protein in washing steps we use pbs with tween but I want to know in PCR ELISA that we work with DNA for washing unbinding bond do we need tweeen or it has negative effect for hybridization?
Since tween-20 is a nonionic detergent it won't really interfere with the DNA binding to the probe, however, you will want it to help disperse the molecules from your reaction tube so that they can bind to the probe. Also, tween-20 helps reduce the nonspecific binding to the plate/well/tube by saturating those possible sites with tween instead. Bottom line, yes, as Netrapal Singh and Karima Al-Salihi have said, you need tween.
For those asking what is PCR-ELISA or wondering if the question is in reference to either PCR or ELISA, PCR-ELISA is basically an ELISA that quantifies DNA/RNA (PCR product) instead of protein or peptide. Streptavidin will bind to Biotin regardless of what Biotin is bound to. So, instead of Biotinylating an antibody, you can Biotinylate a nucleotide sequence (your probe). Adding this to a streptavidin coated plate will result in a DNA probe bound to the plate instead of an antibody, as in a regular ELISA. The rest of it is principally that same as ELISA.
Hope this helps!
-Zack
If you are talking about ELISA, tween 20 or 80 is not needed. Commercial kits come with their wash buffer which has to be re-constituted with distilled water according to the strength (maybe 10X or 20X etc). If you are talking of PCR, most extraction kits do not require tween 20 or 80. After lysis, there are the wash buffers also, which need to be reconstituted by addition of absolute ethanol.
Since tween-20 is a nonionic detergent it won't really interfere with the DNA binding to the probe, however, you will want it to help disperse the molecules from your reaction tube so that they can bind to the probe. Also, tween-20 helps reduce the nonspecific binding to the plate/well/tube by saturating those possible sites with tween instead. Bottom line, yes, as Netrapal Singh and Karima Al-Salihi have said, you need tween.
For those asking what is PCR-ELISA or wondering if the question is in reference to either PCR or ELISA, PCR-ELISA is basically an ELISA that quantifies DNA/RNA (PCR product) instead of protein or peptide. Streptavidin will bind to Biotin regardless of what Biotin is bound to. So, instead of Biotinylating an antibody, you can Biotinylate a nucleotide sequence (your probe). Adding this to a streptavidin coated plate will result in a DNA probe bound to the plate instead of an antibody, as in a regular ELISA. The rest of it is principally that same as ELISA.
Hope this helps!
-Zack
Dear Seyyed Mehdi Mousavi
please read attach article more details about pcr elisa and its application
Dear mehdi mousavi as friend say pcr elisa is a combination method for pcr and elisa for example for detect a special sequence in dna by using probe that is attached to the plate,the steps are similar to elisa with little different .
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