O. Marte

13 Questions 30 Answers 0 Followers

Questions related from O. Marte

Is it normal to have a second peak in chromatograms?

I'm seeing a second call inside most peaks of my DNA sequencing data. This second (smaller) peak is exactly the next base called. Has any one seen this before? Is this a problem with primers? I'm...

17 April 2017 7,441 11 View

How can I get rid of nonspecific band in product?

Hello, I am designing a Taqman assay and have used ABI's Primer Express software to design the primers and probe. I've also had a bioinformatician check that the sequences are in conserved areas...

17 March 2017 4,680 1 View

What effect does low 260/230 have on downstrain applications, like RT-PCR?

There seems to be salt in my RNA after extraction. I proceeded to perform a cleanup step and while the quality (260/280) improved, the 260/230 remains low. I do not have more sample to...

04 May 2016 1,104 5 View

If I add Trizol to whole blood for future RNA extraction, do I have to add heparin or EDTA as well?

Would freezing the blood in Trizol enough?

09 March 2016 8,747 13 View

What are the problems that arise with too much RNA or cDNA in a qPCR reaction?

I'm trying to increase sensitivity (detection) for a gene expression assay. The more RNA or cDNA I add the more detection I get. I need to be able to detect at really low parasitemia, so the...

01 July 2015 3,331 5 View

Can someone explain in some what simple terms why we can't use SYBR Green I master mix with Taqman probes?

03 June 2015 4,753 3 View

Does anyone have experience concentrating RNA using a speedvac?

I've never used a speedvac, but it was suggested to me in order to concentrate my RNA. I was told I need to know which temp and time (and I can't remember what else) to set up as my parameters. My...

27 May 2015 3,504 4 View

Does anyone have a protocol to collect red blood cells?

I have whole blood and want to isolate rbc. I've come across some methods that isolate wbc, but haven't been able to find one for the collection of rbc. Any help would be appreciated.

24 April 2015 7,340 8 View

The first point of a serial dilution to test the efficiency of my primer seems to have inhibitors, what should I do?

I am testing the efficiency of my primers by doing a serial dilution. The efficiency is decent, but when I remove the first point it improves, leading me to think that the first, undiluted point,...

25 March 2015 1,230 4 View

If I decrease the extension temperature in my real time PCR, will I increase the time?

The SYBR Green kit has the extension at 72C for 30 seconds. Should the time change if I decrease the temp?

10 March 2015 4,660 9 View

Is a smear like this in a gDNA gel normal?

I know smear for gDNA is normal, but I don't know what normal is. Can anyone tell me if there is a problem with my samples based on my picture?

06 February 2015 5,036 16 View

How can I run gDNA extracted from rat ear punch on gel?

I need to run a gel before sequencing it. What percent gel should I run it on, and what size ladder is adequate?

30 January 2015 9,663 2 View

Which volume do I use to dissolve my DNA pellet after phenol chloroform clean up?

My starting concentration was 4.8ug, and I'm assuming 50% recovery.

30 January 2015 6,666 2 View