memari

Have you ever tested Qubit 2.0 Fluorometer? and How better is than NanoDrop( Lite)?

And is there any other device and technology for RNA, DNA and Protein quantification?

Which antibody is good for HA-tag? I used santa cruz and cell signaling but both of them give alot of background bands.

Which one is better as reference gene base on these data?
I (New student) have a debate with a PhD student (He has started his PhD three years ago) about choosing a reference gene.
I say that GAPDH is good. But he says that 18s is good.
What is your idea? 1 is non-treated cells and 2,3 and 4 are cells which have been treated differently.

---------------- First Repeat-- Second Repeat-- Third Repeat-- Average
1-- ACTIN-  14.0449068-- 14.0022355-- 13.9916405-- 14.0129276
2-- ACTIN-  14.0670855-- 14.0977196-- 13.9009442-- 14.02191643
3-- ACTIN  13.9944372-- 13.7456839-- 13.7724482-- 13.8375231
4-- ACTIN- 14.2457309-- 14.2877965-- 14.0749623-- 14.2028299
1-- 18S-   9.2736629-- 7.4010807-- 8.5428005-- 8.405848033
2-- 18S-   9.2615854-- 6.7311396-- 6.7095866-- 7.5674372
3-- 18S-   9.2900732-- 9.30192-- 8.5359895-- 9.0426609
4-- 18S-   8.5324967-- 8.5267837-- 6.9371083-- 7.998796233
1-- GAPDH- 17.3842867-- 17.5144233-- 17.2672231-- 17.38864437
2-- GAPDH- 16.9393409-- 16.981744-- 17.1600726 17.0270525
3-- GAPDH- 17.477777-- 17.2306059-- 17.3061118 17.3381649
4-- GAPDH- 17.3204051-- 17.0312215-- 17.1166215-- 17.1560827
1-- ACTIN-2- 15.6542578-- 16.0506853-- 15.308673-- 15.67120537
2-- ACTIN-2- 15.4098156-- 15.4334925-- 15.0512194-- 15.29817583
3-- ACTIN-2- 15.3326059-- 15.3368817-- 15.2771232-- 15.31553693
4-- ACTIN-2- 15.6690638-- 15.697236-- 15.5794577-- 15.64858583


I have another question:
Do you know how many differences between CT values of three repeats of a reference gene are acceptable?

I wonder why authurs in journals recommend pre-blocking dynabeads by BSA, while the Company of Invitrogen says that "BSA blocking may not be very successful, as surface adsorption is not promoted between the hydrophilic bead surface and the hydrophobic BSA."

https://www.invitrog...ion-FAQs.html#7


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Babak

I wonder why some biologists use PIPES pH 8.0 in lysis buffers in Protocols?
whereas the buffering range of it is 6.1-7.5 pH.

You can find those protocols even in high rank universities.