What are the best ways to reduce generation of chimeric molecules during PCR?

The chimeric molecules generated during PCR are really bad for our study on recombination, since the chimeras look exactly like recombination. I wonder what and how (would be better if protocol is...

06 July 2016 9,547 5 View

What is the average disance from a mismatched base for a base substitution occurring during mismatch repair?

Especially in mammalian cells.

05 June 2016 1,312 1 View

What percent of template can be synthesized during PCR?

For example, the parameters for PCR are denaturing at 95, annealing at 58 and elongation at 72, while the melting temperatures of primers are around 60.

04 May 2015 4,344 13 View

Does anyone have experience about the efficiency for linear DNA synthesis?

We are trying to synthesize mouse DNA using "linear" synthesis in very limited number of cycles without "denaturing". Do you have any experience for linear DNA synthesis, especially about the the...

11 December 2014 5,867 0 View

Which human genes are strongly suppressed by wild-type p53 in human cells?

I am interested in looking for those genes that are expressed more than five fold higher when p53 is lost or mutated.

11 December 2014 9,495 2 View

Which PCR product clean-up kit you would like to recommend?

I am trying to clean the PCR products for deep sequencing, and I would like to know which kit of which company you think is the best (based on your experience if you have done). Thanks.

11 December 2014 2,965 7 View

How likely DNA double strands are seperated (melted) at lower than melting temperature?

For example, how many percent (if any) of DNA is melted at 5 degree lower than the melting temperature? Thanks.

11 December 2014 578 8 View

Do you know of any high-yield genomic DNA extraction kit/protocol?

11 December 2014 945 3 View

Is there a way to reduce the interference from genomic DNA on your template-primer pairing?

For example, whether long DNA fragments or shorter ones (such as with restriction enzymes) interfere with the pairing of the primers to the templates of interest with different efficiency or not.

11 December 2014 2,314 2 View

Is digestion of genomic DNA helpful to increase the pairing of primer to a specific genomic region of interest?

We are trying to test the efficiency of pairing of a primer to a region of mouse genome. We are thinking of digesting genomic DNA before pairing and wonder whether the longer or the shorter...

11 December 2014 6,149 2 View

How can I measure MMP-1 protein activity or protein level from liver tissue?

Instead of checking only the mRNA level, I want to check the active protein level of MMP-1 in Liver tissue from mice. How can I do that?

03 March 2021 1,763 2 View

Does any one know how i can prepare liver tissue with transmission electron microscope with good fixation?

i have problem in preparation of liver tissue with bad cell organelles and how i can adjust osmolarity

03 March 2021 8,890 1 View

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 2 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

B-mercaptoethanol (BME) protocol?

Hi everybody, I have been running a multiplex IHC which allow me to stain the same tissue with different markers (Primary Ab + Secondary conjugated Ab) (AF487, AF555, AF647). Since I would like to...

27 February 2021 5,028 2 View

Looking for genomic DNA extraction kits from mouse tissue (centrifuge-based, not spin-column)?

Hello. I am looking to extract genomic DNA from mouse brain and lungs and I need a kit that is not spin-column based because I need to scale up the solution amounts for 400mg tissue (most cap at...

25 February 2021 155 2 View

Has anyone tried to scan fluorescence IHC slides with microarray scanner?

I have an old slide microarray scanner (VIDAR Revolution 4200) and would like to give it a "second life", as IHC scanner array (fluorescence dyes), FISH over tissue or similar. Has anyone tried...

24 February 2021 9,210 1 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

How to improve RNA isolation?

Hi, I have been struggling with a weird issue for my RNA samples. I used the Qiagen RNA plus mini kit. For some reason no matter how careful I am with the washes, when I measure the RNA, I get...

22 February 2021 1,286 6 View

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22 February 2021 9,932 11 View