Chelex vs. Glassmilk extraction approach for DNA sequencing?

Hi, I am looking into methods to extract DNA from fish tissue. Our lab has used the glassmilk approach in the past but recently switched to using Chelex. I was wondering if anyone knew of any...

24 September 2019 7,467 1 View

Is it possible to find the average of responses to a self-report scale?

My participants have rated themselves 1-10 on a series of 19 statements. Those statements are supposed to represent 5 different constructs. So for example, questions 2, 6 and 11 are all about...

06 February 2019 3,473 2 View

How long can PCR products labelled with FAM dye be stored before it begins to degrade?

I have PCR products stored at -20 degrees (Celsius) that are tagged with FAM dye. How long can these be kept before the fluorescent signal diminishes?

01 December 2018 2,081 3 View

Low signal - Circadian luciferase assay in cell culture?

Hi, we are running circadian luciferase assays in cell lines (u2os and 3t3) driven by two different circadian promoters. We're reading the assay in a TECAN m200 luminometer using a black clear...

01 November 2018 7,642 1 View

How to calculate protein interaction energy using VMD NAMD2?

I am using the VMD plugin QwikMD to do MD simulations of my protein-protein complexes. Hereafter I am using the NAMD2 analysis to calculate my total energy of the system along with all of the...

11 May 2018 1,481 5 View

What statistics should I used for Likert Scale?

Was wondering if anyone could help me, I am in my third year and I'm in dissertation hell. I have used 5 point Likert Scales in my questionnaire (Strongly agree, agree, so on). I'm now looking...

19 January 2018 6,122 5 View

Liquid culture colony PCR technique?

I need to do a pcr on a liquid culture of E.coli. I'm amplifying a 150bp region of gyrA, using primers that I designed. I'm using NEB Next Ultra 2 Q5 MM. For the reaction, I am using 5ul NEB, 0.5...

25 October 2017 5,995 5 View

Another question on transcription and coding, can anyone provide up to date help?

Hi everybody, I'm undertaking my first qualitative research and am somewhat overwhelmed by the (general consensus) of time it will take to transcribe qualitative interviews - I am intending on...

10 October 2017 3,011 3 View

I would like to store 10 uL aliquots of bacterial culture. What would be the best way?

I need to store 10ul aliquots of bacterial culture , would the best way be to put them in PCR tubes with some gylcerol (10ul 50%) at -80?

31 January 2017 2,848 3 View

What distinguishes the fourth industrial revolution from previous radical economic development eras ?

04 March 2016 6,069 11 View

Can bacterial suspension be pipetted directly onto petri dish and agar poured onto it?

Hi, recently I encountered a protocol for plating which was not the usual method I have been practicing (pour LB agar onto plate, allow it to solidify, pipette bacterial suspension onto plate,...

28 February 2021 7,292 3 View

Why does my MTT Assay remain in purple color and does not change to yellow?

19 January 2021 5,098 4 View

Is the pipetting order of the positive and negative controls in the PCR important to avoid in a possible contamination?

I have been watching bands form on my negative controls. I have already changed all reagents, optimized the hygiene of materials and the environment and increased care in pipetting.

07 December 2020 9,090 3 View

Should I vortex during qPCR solution mixing?

To do qPCR we have to mix solution or do serial dilution. Like, 1. cDNA dilution 2. Primer dilution 3. mixing SYBR Green, cDNA and Primer in PCR tube. My question is for doing this mixing when...

25 October 2020 2,576 11 View

Why my absorbance values are in negative for antioxidant assay by dpph method ?

plant extract stock is 10 mg/mL from which I pipetted 20, 40, 60, 80, 100, 200 uL and made up to 3 mL using methanol then i added 1 ml of 0.004% dpph solution (dissolved in methanol) incubated in...

02 September 2020 1,218 2 View

How can I digest the brain tissue in healthy mice as evaluating the apoptosis using flow cytometery by annexin V/propidium iodide (PI)?

We digested the brain tissue with Trypsin ( 1x) for 15 min (all steps were performed in cold conditions) and suspend the cells by pipetting up and down (3-4 times) to the single-cell suspension....

22 August 2020 5,611 4 View

Culturing CHO-freestyle cells without shaking?

Hi everyone! Recently I started working with CHO-Freestyle suspension cells, but our laboratory does not have an orbital shaker. I tought it would be okay without it, so I just used a normal T25...

20 July 2020 4,597 1 View

How to improve the lentivirus transduction efficiency in jurkat or parimary T cells?

Hi, I am transducing jurkat cells with lenti-virus but my efficiency rate is low... my transducing method is this: 1. I defreez viruses and put them 20 min in room temp. , 2. resuspending them in...

25 June 2020 9,077 3 View

No title

I have tried transforming E. coli BL21(DE3) cells with a heat-sensitive plasmid using electroporation. The plasmid contains an ampicillin resistance cassette. After streaking the cells on an LB...

28 April 2020 5,540 6 View

Cutting the plasmid pGFPuv with enzymes?

At which positions these enzymes cut the plasmid pGFPuv.? BamHI, NdeI how to Calculate how many fragments of which sizes you can expect after each of the reactions.? what is the pipetting...

01 February 2020 3,998 3 View