Degenerated primers for sequencing?

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Does anyone have a sequence for the human CD34 promoter?

I'm looking for the human CD34 promoter sequence to engineer into a vector construct. Can anyone recommend a good source for this sequence? Thanks very much, Laura

25 February 2021 8,205 1 View

Quantifying leaking DNA from bacteria - what's the secret?

Hi, all! I need to determine if my treatment causes membrane damage in bacteria (S. aureus and E. cloacae), but I cannot use a fluorescent probe such as PI and similar ones because my compound is...

24 February 2021 2,006 2 View

Is anyone else having a similar situation?

I'm looking for the genetic sequence for the TAR region of HIV. If I put Transactivator Response Region, and HIV, and "genetic sequence: into a browser...is it not unreasonable to expect to be...

10 February 2021 658 5 View

Can a mRNA be degraded but still observe a protein over-expression?

Hello there, I'm currently trying to over-express a gene by transfecting the cells with a pcDNA6 plasmid containing my target gene. When I measure the mRNA for my gene of interest, the levels are...

03 February 2021 4,112 4 View

Are there any solutions for remote eye-tracking?

Since with the pandemic it doesn't look like we are going back to the lab soon, in my lab we were looking for solutions to conduct remote webcam-based eye-tracking. I've been looking at...

31 January 2021 7,315 5 View

Sample Size for Quasi-Experimental Quants using one-tailed t-test?

Hi, sorry I'm stumped. I'm going to do a study on the leadership styles (using the MLQ Form 5X) of minority women administrators in medical schools. How will I calculate the sample size? Thank...

30 January 2021 2,865 4 View

I'm curious whether there is any research available on the use of AI related to diversity&inclusion (efforts)?

I'm looking for studies or conceptual papers on the (potential) uses of AI for enhancing diversity and inclusion within organization. The biases concerning the use of (big) data are known and...

18 January 2021 4,222 4 View

Where can I purchase NanoLC emitter fused silica picoTip?

Hi, I'm working with NanoLC/MS for both Metabolomics and Proteomics. We usually make our nano columns using emitters from NewObjective. Unfortunately New Objective, due to the pandemic, is now...

10 January 2021 1,176 4 View

What can be the reason of having high Cq-values for reference genes?

Hi, Once again I am facing an annoying problem. This time in my barley qPCR, I am trying to detect a virus (BYDV) in plant samples. I get rather high Cq-values for 2 of the 3 reference genes...

06 January 2021 4,685 9 View

Thawing or defrosting methods for frozen soil samples?

I have soil samples that have been frozen for a while and I need to re-acclimate them for a series of analyses that require incubation at 25C. I have been looking for laboratory methods but seem...

04 January 2021 4,862 3 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 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

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 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

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

Genescan gives 3 peaks instead of one?

Hi all, I am doing a genescan analysis for a deleted exon in the patient sample. The amplified region is 215 bp. In the patient sample because of homozygous deletion, no peak is observed while in...

23 February 2021 5,645 1 View

QPCR reaction efficiency with odd results what does this indicate?

I am working on a CFX 96 machine to determine reaction efficiency of my gene of interest. After performing a serial dilution of my cDNA template, I was able to find great reaction efficiencies for...

23 February 2021 809 3 View