Do all the mRNAs of pox virus are polyadenylated at 3' end?

Hello everyone, In eukaryotic cells, all the mRNAS are polyadenylated at 3' end. I want to ask whether this is the same case for pox viruses also. Do all their mRNAs are polyadenylated at 3'end...

04 May 2019 3,219 2 View

Can we use freezed nuclear lysate forDNMT activity analysis from cells?

Hello everyone. I want to do DNMT activity analysis by using prostate cancer cells. I want to know whether we can store the nuclear lysate for 2-3 days in -20 degree before performing activity...

05 June 2018 6,439 7 View

Do we need to quantify bisulfite treated DNA for using it as a template for methylation specific PCR?

Hello everyone I have bisulphite converted my DNA using kit from Zymo research now I have to do methylation specific PCR. I have to check effect of my drug on methylation status of diff. genes...

10 November 2017 8,291 4 View

Can we use autoclaved DEPC milliQ water for bisulpgite conversion and methylation specific pcr?

I am going to do ms pcr but I have doubt on water milliQ water because previously RT pcr result was not coming by using that water. So I borrowed water from another department but that water was...

09 October 2017 3,899 4 View

RBC contamination in WBC pellet during DNA isolation from blood?

Hello everyone, I isolated DNA from freshly taken Human blood but during processing I was getting RBC contamination in pellet .I washed with RBC lysis buffer 6-7 times followed by...

09 October 2017 5,351 3 View

DNMT activity assay?

06 July 2016 2,163 5 View

Has anyone done RT PCR for Cyclin D2 , p21 and p27 in DU145 cells ?

I have done RT -PCR for Cyclin D2, p21 and p27 in du145 , but did not get any band . in p21 , there was v. faint band but when I repeated the same , I did not get any band for p27. does these 3...

03 April 2016 839 1 View

Why do we need to do the conditioning of vessels with media or PBs in rotary cell culture system?

31 December 2015 4,391 2 View

How to freeze LNCaP cells?

What is the best way to freeze LNCaP cells? ATCC recommends complete medium plus 5% DMSO. some people use 10% DMSO, 20% FBS and rest complete medium. Some use 90% FBS and 10% DMSO. Please tell me...

31 December 2015 2,323 3 View

What is the difference between LNCaP cells and LNCap FGC cells. Can we use LNCaP FGC cells in place of LNCaP cells?

I have to use LNCaP cells for Methylation specific PCR, but LNcaP cells are not available but LNCaP FGC cells are available. So Can I use these cells in place of LNCaP cells?

10 November 2015 6,498 3 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

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 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

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

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

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

01 March 2021 5,311 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 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