Can anti-6xHis antibody detect 8xHisTag?

Hi researchers, As the title shows, recently I made a construct with a 8xHis tag at N ternimal of HBx (154 aa). The voctor is pCDNA3.1+ and the sequencing result showed everything was correct. I...

10 November 2017 6,357 5 View

Anyone use this PCR mycoplasma detection protocol and get a 100bp product?

Hi researchers, I'm using a PCR mycoplasma detection protocol which is found online and it works well for me. Link as...

09 October 2017 4,263 4 View

What is the difference between I-TASSER and QUARK protein structure prediction?

07 August 2017 2,179 2 View

Why cannot detect protein after over-expressing by lenti-virus?

Hi researchers, I want to over express a protein in Hep3B cell line. The expressing vector I use is attached here. The packaging vectors are pMD2.G, pRSV-Rev and pMDLg/pRRE. The amount I use for...

06 July 2017 1,628 4 View

Is Swiss prot ID unique for each transcript?

03 April 2017 2,260 3 View

Secretory protein with Flag at N-terminal?

Hi researchers, I made a construct with a Flag tagged to the N-terminal of my aim protein. I've already done the lentivirus infection and selected for over 2 weeks. I did a real time PCR to check...

31 December 2016 6,854 7 View

What is the size for BMP2 protein in western blot?

31 December 2016 7,301 4 View

Which part of the siRNA matters most?

Hi all, I want to order one siRNA from one publication. In the publication, they used 3 siRNAs and I want to order the most efficient one first. This is a 19nt siRNA. But when I do the blast,...

09 October 2016 381 4 View

What does XM_xxxxxxxxxx mean in NCBI?

Hi researchers, When I do primer blast, the result will show me an ID of the gene that my primer will bind to. If I'm not wrong, NM_xxxxxxxxxx means it's a human sequence. Then what does...

08 September 2016 4,079 2 View

For secreted protein, can we do western blot using cell lysiate instead of ELISA of cell culture supernatant?

I want check the protein level of BMP2 (Bone morphogenetic protein 2). I see someone use cell lysiate to do western blot while others use cell culture supernantant to do ELISA for BMP2. I know...

08 September 2016 1,510 4 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 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

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 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

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

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

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