Is it possible to do a No Reverse Transcriptase control with the Luna One Step qRT-PCR Kit?

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Absorption coefficient of atmosphere at sea level versus wavelength?

I'm looking for some raw data for the transmission through the air in my lab, specifically in the MIR region. I've found a lot of data online that gives transmission vs wavelength, but the units...

22 December 2020 9,039 2 View

How to test if two subsets of a distribution have different correlations/regressions with other variables?

I have numerous non-normal distributions that, for the most part, have the same shape (n = 190). The variables are test scores and have a floor value of 5 and maximum of 50. They appear to be...

22 October 2020 9,305 3 View

Plant checklists for Borneo and Sumatra?

Dear RG Community, Can someone please give me references to publications (or online websites) that include plant checklists (vouchered if possible) for Borneo and Sumatra? Thanks in advance, Zach

20 September 2020 6,202 4 View

Should I use square transformed mulitple regression or ANOVA?

I am building a model with: - Non-normal continuous dependent variable (a score on a questionniare which does not have literal clinical meaning) - it appears fairly normal on histogram but there...

13 August 2020 3,439 9 View

Adding a transcriptional repressor before a promoter sequence in my plasmid?

I am performing a KI experiment using CRIPR/Cas. Our 5' homology arm is the 5'UTR of the gene of interest and therefore contains the endogenous promoter. I am looking for a repressor sequence to...

19 May 2020 5,088 3 View

How to design a vector that uses loxP for removal of blasticidin resistance gene?

Using CRISPR/Cas9 and homoly directed repair I have knocked in a degron tag onto the start of a protein of interest in 293T cells. WB has confirmed that ~5% of protein alleles have recieved the...

17 March 2020 4,470 1 View

A heterogeneous fluorescence sensor appears to exhibit second order scattering. The "scattering" responds linearly to target analyte, is this normal?

I am performing experiments on a suspension of fluorescent material that is quenched in the presence of a target analyte. I excite at 279nm and see a peak around 560nm. When I add analyte solution...

10 February 2020 2,743 4 View

What are the techniques to validate qualitative research instruments?

I am currently doing a quasi-experimental research. I have decided to use both qualitative and quantitive methods in collecting the data. For the qualitative data, I have an observation card and...

08 October 2019 4,007 5 View

How a researcher can validate his upcoming research instruments?

I will be conducting an action research in one of the schools at my place of working and my supervisor has asked me to validate my instruments before I can use them during the research process. I...

11 December 2018 1,355 3 View

Bizzare qRT-PCR amplification curve - Why does signal drop to zero near end of run?

Sorry for the poor image quality, I am working with a fairly old real-time PCR system (Bio-Rad MyIQ on iQ 5 version 2.0 software). I am using primers, cDNA, and supermix (Bio-Rad iQ SYBR Green)...

12 August 2018 3,553 4 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

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 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

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View