What are the odds of dragging viable phage from PCR reaction (was template) and column DNA purification?

09 January 2017 1 1K Report

we used cDNA library (linear in phage state before plasmid closing) as a template for PCR mediated addition of adaptors..

will the phage survive multiple cycles of boiling? If yes then what can I do other than the standard DNA purification to assure No phage dragging?

Gil Zimran

Thank you for the helpful answer!

Badges
Science topic

More Gil Zimran's questions See All

How can I improve my electrophoresis technique?

Hello. I'm making non-denaturing PAGE of DNA fragments for analysis of polymorphisms, but I'm getting wavy bands. If somebody can help me, I will be very greatful. First of all, here's my recipe...

15 March 2024 7,699 1 View

Can cy5 be fixed using formaldehyde ?

Hello I am planning to deliver a cy5-labeled drug to mice, and then check its distribution in different organs. In case I wish to preserve the tissues for later research, will fixation (using 4%...

12 November 2023 4,958 1 View

Lentiviral transduced cells loose insert?

I have infected iPSCs with a lentivirus and sorted GFP positive cells. But after just a few rounds I am finding cells without GFP in culture. Had anyone seen this? Maybe cells are segregating...

17 October 2023 1,611 1 View

SORTING of iPSCs cells?

We are trying to purify human iPSCs using lentiviral induced GFP expression. But is being really difficult to achive this. Cells are usually dying after sorting. Any experience on this? anyone...

25 September 2023 6,985 2 View

In modern shunt system what is a typical distal catheter length within the peritoneal cavity?

In the 2010 era, it was typical to provide distal catheter lengths that were 2 to 3 times the height of the cavity to allow it to slide and bend away from obstructions. The force necessary to stop...

02 September 2023 10,029 0 View

Non-parametric Kruskal Wallis instead of a two-way ANOVA?

Can I apply a non-parametric Kruskal Wallis test instead of a two-way ANOVA when assumptions of normality and heterocedasticity are not met?

11 April 2023 4,229 11 View

Why is it expected to get a high percentage (≥80) of uniquely aligned reads in RNA sequencing?

I started learning how to use Tophat tool. I used it to check RNA sequencing and I calculated 64.91% in the percentage of uniquely aligned reads. I read in article that in RNA sequencing we should...

14 November 2022 7,252 3 View

Does anyone know about transformation methods in Mycetohabitans spp?

I'm having trouble transforming my bacteria.

09 August 2022 2,357 0 View

A question regarding the ultracentrifugation method for exosomes isolation?

Hi, In the ultracentrifugation method, there are several steps in which one collects the supernatant ,discard the tube with the sediment, transfer the supernatant into a new tube for a higher RCF...

14 November 2021 182 3 View

Who has already compared sediment cores downstream of dams with proxies of discharges time series and/or suspended sediment fluxes?

The objective is to assess the impact of dams on the reduction of solid transport.

31 May 2021 3,152 0 View

How to improve plaques in Double Layer Agar Assay?

Hi all! My thesis groupmates and I are working on purifying an E. coli bacteriophage. The phage was handed down to us by the previous thesis group, but they only managed to purify it to about...

06 August 2024 5,801 4 View

Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...

06 August 2024 1,495 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 View

The Curse of Evolution and Complexity?

Brain and body mass together are positively correlated with lifespan (Hofman 1993). The duration of neural development is one of the best predictors of brain size, and conception is the best...

05 August 2024 6,247 3 View

For an in-vitro drug release study, what molecular weight cut-off (MWCO) dialysis bag is required for a 117 kDa protein?

kindly reply me. Thanking you in advance.

05 August 2024 7,727 4 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View

How to start a Molecular Dynamics Simulation?

Is it possible to conduct a molecular dynamics simulation to see the effects of a specific carbohydrate on the structure of lipids (e.g., micelle structure)? I am a beginner in this field and plan...

03 August 2024 3,371 3 View

Which will be the best software for the Hydration shell analysis with molecular dynamics?

I am using a windows system, what software I should use for hydration shell analysis with molecular dynamics?

02 August 2024 3,143 4 View

Can anyone provide me with molecular docking softwares/ websites?

Molecular docking software/ websites?

02 August 2024 8,704 7 View

Where to find a gene list for CRISPRa/i library screening of regulatory factors that affect pathogenic Th17 differentiation in PBMC?

I want to perform a CRISPRa/i library screening for candidate regulatory factors that affect pathogenic Th17 differentiation in PBMC. But I am wondering how to define the size of gene library, so...

31 July 2024 2,150 0 View