How to get going smRNA FISH on fresh frozen breast tissue?

Hello, I am having difficulties to get smRNA FISH going on fresh frozen breast tissue. The technique worked well on MCF7 cells. We tried the same probes on tissue sample, and we haven't had...

05 June 2019 7,841 0 View

How can one ensure ligation of a 300bp fragment into 5369bp host?

09 October 2014 3,484 16 View

How to troubleshoot Site Directed Mutagenesis?

I did SDM with double primers PCR. This was followed by dpn1 digestion.The gels showed amplicons but when I transformed there were no colonies. The polymerase used was pfu and Tm was 60 degrees...

31 December 2012 1,081 17 View

Docking

I would like to dock an enzyme with its substrate. Can anyone suggest any free software which I can use and which has a tutorial provided?

06 July 2012 6,756 22 View

How to perform MSA for two domains of a single enzyme by clustal X?

I have been trying to do clustalx for my sequences of Polyketide synthase. Many of my sequences have two Acyl Carrier protein (ACP) domains. The problem is that clustal x is not accepting the...

05 June 2012 6,283 17 View

Fungal DNA isolation

I am not getting my results for DNA isolation from Aspergillus. Can it be because the lysis buffer did not have SDS?

05 June 2012 1,097 7 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

Does anyone know an alternative product for cardiac Troponin T antibody for Immunofluorescence (Thermo # MA512960)?

We have been using Thermo Fisher anti-cardiac Troponin T antibody for immunofluorescence staining in FFPE mouse heart sections but it was recently discontinued. The literature shows it to be the...

01 March 2021 4,937 1 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 3 View

No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Markers of mitochondrial activity in FFPE tissues?

Hello everybody, Does someone have any experience with assessing/extimating mitochondrial activity on FFPE tissues? Precisely, no TOM20, ATP5B, etc: we are trying to get an approximative idea of...

24 February 2021 4,618 4 View

Errors in DNA sequences?

I am trying to better understand the scope of DNA replication and sequencing errors, e.g. 1. I have seen similar error rates of 10e4 to 10e5 for cell & instrumental DNA replication,...

24 February 2021 4,397 3 View