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Discussion

5 years ago

Daniel McKenzie

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

5 years ago

R. Verma

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

Discussion

4 years ago

Paul G. Macon

4

Votes

2

Comments

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Experiment: RNA isolation / purification Tissue

Discussion

4 years ago

Milena Alexeyeva

3

Votes

2

Comments

DNA insert using CRISPR

I would like to excise a large strand of DNA and insert a new one using CRISPR. My problem is that my strand will be a little over 1kb and I am not sure if this is going to be a limiting factor. Also, how long should the homology arms be for a region of this size?

Experiment: CRISPR Human

Discussion

5 years ago

M. Daecher

2

Votes

2

Comments

Live/dead assay Bacteria

Hello everyone! I am going to do a live/dead assay for my cells and I saw that I can use both fluorescence and absorbance as my detection method. Is there a difference in the results depending on the method? Is one method preferred over the other in certain situations?

Experiment: Live / Dead assay bacteria

Discussion

5 years ago

Jody Hancock

2

Votes

2

Comments

How can I preserve leukocytes for DNA methylation profiling?

I would like to preserve leukocytes for future epigenetic analysis. How can I preserve them effectively in order to perform DNA methylation profiling at a later time?

Experiment: DNA methylation profiling Gene specific profiling

Discussion

5 years ago

Aaron Stege

4

Votes

2

Comments

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

Experiment: RNA isolation / purification Tissue

Discussion

5 years ago

Israel Lev

2

Votes

2

Comments

How can I improve my DNA yield?

The DNA concentration after using this DNA isolation kit is sometimes too low and thus it is not sufficient for my follow-up experiments. How can I improve it?

Experiment: DNA isolation / purification Bacteria

Discussion

5 years ago

Stefan Fuhrmann

1

Vote

2

Comments

Will presence of EDTA effect cDNA synthesis

I intend to use iScript cDNA Synthesis Kit in order to synthesize cDNA for qPCR. I have confirmed that my RNA is pure however, according to my extraction protocol I have suspended the RNA in TE buffer containing 1mM EDTA. Will the presence of EDTA have an effect on cDNA synthesis?

Experiment: cDNA synthesis Bacteria

Discussion

5 years ago

A.C.Burton

1

Vote

1

Comment

Multiple gene silencing using ShRNA

Hello, can someone here help me? I am trying to silence e-selectin and ICAM-1 in endothelial cells. I would like to know if this is possible using shRNA

Experiment: shRNA gene silencing Mouse

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