Discussion

1 year ago

Eufrosina Sagese Italy

3

Votes

3

Comments

Is a knockdown using shRNA permanent?

Is a knockdown using shRNA permanent and if not is there a known duration?

Experiment: shRNA gene silencing Human

Discussion

1 year ago

Jana Fassbinder Germany

3

Votes

1

Comment

What is the optimal concentration for primers in qPCR?

What is the optimal concentration for primers in qPCR? My total volume is 20μl per reaction.

Experiment: PCR Quantitative real-time PCR

Discussion

1 year ago

Nicolai Brandt Denmark

3

Votes

2

Comments

What DNA isolation kit would work for insect samples?

Hello everyone! I am currently using different DNA isolation kits to extract DNA from insects. Even though I am able to successfully extract DNA I would like to maximize the yield. Do you have any tips that might help me with that even if the kits are not specifically designed for insect samples?

Experiment: DNA isolation / purification Tissue

Discussion

1 year ago

Paul G. Macon United States

2

Votes

4

Comments

Some help with RNA isolation using Trizol

Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?

Experiment: RNA isolation Bacteria

Discussion

1 year ago

Sander J. Mikkelsen Denmark

2

Votes

2

Comments

endothelial cells vs HUVEC cells?

What is the difference between endothelial cells and HUVEC cells for angiogenesis assay?

Experiment: Angiogenesis assay human

Discussion

11 months ago

S Janardhan Belgium

3

Votes

2

Comments

Reactivate dormant Lactobacillus Paracasei

Hello fellow Labettors, I would like to know what would be the best method/medium to reactivate my dormant Lactobacillus paracasei cells?. Any suggestions are greatly appreciated.

Experiment: Bacterial cell culture media Lactobacillus paracasei

Discussion

7 months ago

Mario Udinese Italy

3

Votes

2

Comments

Floxing mice with CRISPR

Hi everyone! I am planning on floxing mice with CRISPR but I am having trouble deciding which region to target. Do you have any tips on choosing?

Experiment: CRISPR Mouse

Discussion

1 year ago

Alex Santos Ribeiro Brazil

3

Votes

2

Comments

cDNA conversion with low concentrations

I have extracted RNA from brain tissue but my RNA concentrations are as low as 5ng/ml with my highest being around 80ng/ml. Will I be able to perform cDNA conversion with concentrations as low as these?

Experiment: cDNA synthesis Tissue

Discussion

1 year ago

Daniel McKenzie United Kingdom

2

Votes

2

Comments

How “strong” is the PCR product of methylation specific PCR?

Does anyone know how “strong” the PCR product of methylation specific PCR is? I kept my PCR products at 4C for about 3 weeks and then at room temperature for another week. Will I be able to use them for sequencing?

Experiment: PCR Methylation specific PCR

Discussion

11 months ago

R. Verma India

2

Votes

2

Comments

Multiplex PCR and qPCR

I have been doing multiplex PCR and qPCR separately. Is there a way to combine them in a single reaction using Sybr Green?

Experiment: PCR Multiplex PCR

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