Esteban Barila

7 Questions 13 Answers 0 Followers

Questions related from Esteban Barila

Sticking Brain slices on Superfrost slides?

I'm not familiar with brain IHC, but I'm experienced in fixing (with PFA), crioprotecting with sucrose solutions and embedding in OCT some other tissues.I've reacently tried a whole mouse brain...

30 April 2018 1,353 3 View

What protocol would you recommend for membrane protein extraction from tissue?

I need to enrich the membrane proteins from a tissue within the eye of a cow. I've done total homogenates without success. Which protocol would you recommend?

18 August 2017 9,132 1 View

DNAse digestion of RNA samples?

I'm extracting RNA with a trizol-like reagent called Transzol. It looks like there's variable amounts of gDNA on a high molecular weight band on the agarose gel right above the ribosomal...

17 January 2017 573 0 View

Could some mRNA's not get fully reverse transcripted?

I'm relatively new to Reverse transcription. I have worked with cDNA before but dealing with RNA extraction and reverse transcription is new for me so bare that in mind. I'm studying the...

27 December 2016 729 2 View

Low RNA yield, what am I doing wrong? (SOLVED)

I just did my first RNA extraction from tissue using a trizol-like reagent called "Tranzol". I must have eluted it to 70ul final volume but I'm having poor yields of approximately 150ng/ul, low...

31 October 2016 6,586 5 View

In dealing with primers with very different Tm's, how can I work around the PCR?

The forward primer has a tm of about 60ºC and the reverse, which is the Sp6 promoter primer, is merely 45ºC. Last couple of PCRs where getting better as I lowered the temperature of annealing to...

30 May 2015 8,927 5 View

PCR overlap extension with a small 29bp overlapping region (Tm=67ºC) Should I worry a 72ºC extension step?

I'm doing mutagenesis on a plasmid with mutagenic primers by lifting two overlapping semgents carrying a single point mutation (this is not the stratagene method). I will use these two sequences...

01 January 1970 1,321 14 View