Any advice on how to isolate DNA/RNA from mouse sperm?

More Tian Chi's questions See All

T lymphocyte apoptosis in mice

We have found that deleting a transcription factor in Tregs causes fatal autoimmune disorder in mice, concomitant with accumulation of effector memory CD4 cells (CD44hi CD62L low) in blood, LN and...

06 July 2017 7,136 1 View

Why is FACS analysis of CD81 problematic?

Hi we are examining CD81 expression in B cells, macrophages and neutrophils in PBL from mice. To this end, we collected 10 ul blood, lysed RBC, and stained in 10 ul antibody cocktail (B220, CD81,...

07 August 2016 6,179 3 View

Problem with bicistronic expression mediated by P2A self-cleaving peptide

Hi, I have a construct that expresses a certain gene (1.7 kb) together with the downstream GFP; the co-expression is mediated by P2A. In transfection assays, GFP expression was very bright....

01 February 2016 9,489 5 View

Is Tet-controlled binary transcription regulatory system applicable to C. elegans?

Tet-controlled binary transcription regulatory system is widely used in mammalian cells. Does anyone know if it is applicable to C elegans (in vivo and in cell lines)? If not, why? Thanks Tian

11 December 2015 4,082 3 View

Has anyone used the PCR machines and gel documentation systems made by Labnet?

These equipments from Labnet look very enticing, but oddly, seem very unpopular, as none of my colleagues knows about them, and I could not find any customer reviews. Any feedback highly appreciated.

05 June 2015 6,526 4 View

Binary Logistic Generalized Linear Mixed model in SPSS. Is it doable?

I want to analyses the proportion of swimming sperm of three species of fish in two salinities. To analyse the proportion of swimming sperm in a Generalized Linear Model, I would use a Binary...

03 March 2021 2,297 3 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

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

How to repeat an operation on an atom selection for multiple times in PyMol?

Hi, I am trying to construct a multi-layer fibril structure from a single layer in PyMol by translating the layer along the fibril axis. For now, I am able to use the Translate command in PyMol...

02 March 2021 4,569 4 View

What analysis to choose with a large neste dataset and clearly skewed or kurtotic distributions?

Hello! I have a dataset of n=3000 nested within 8 countries with approximately 200 or 400 responses in each country. I originally planned to perform multilevel modelling with 4 dependent variables...

02 March 2021 6,865 1 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

Is there a method of determining feed digestibility without the need to re-pellet the feed?

I am aiming to determine the ADC values of a commercial fish feed, but I do not have a steam pelleter or extruder available to me. Ideally, the pellets could be coated with some inert marker....

28 February 2021 7,967 3 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

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

Tian Chi

Thanks Chris. You understood it right, but I am afraid the sperm RNA content too low for Trizol. Qiagen has a "Allprep" kit for simultaneous DNA/RNA purification, but that is not optimized for sperm, unfortunately.

Vincent Coulon

I agree with Christoph, except that I think DNA is in the organic phase in acidic phenol (and proteins at the interphase).

If it is simply a matter of amount, you can add glycogen, tRNA, or acrylamide as a carrier and still use TRIZOL (small volume to increase concentration). Longer centrifugation times may also help.

Marijo Greene Kent-First

I have used Promega products to isolate sperm DNA and RNA. Modifications of their Wizard system works great as does Magnasil.

Sumira Phatak

I have been using TRIzol for DNA/RNA isolation and am experiencing the same issue with poor DNA concentration and purity. Did you ever find a solution?