Which Do You Prefer: Kit or Freeze/Squeeze?

I am trying to extract a small (

01 February 2019 8,329 1 View

Zero Blunt Cloning - Where do these bands come from???

After PCR using Q5 pol, I ligate the insert into pCR-Blunt using T4 ligase. Transformation into DH10 is successful and a few colonies are mini-prepped. The plasmid is then cut with EcoR1-HF and...

10 January 2018 2,508 8 View

Zero Blunt Cloning - Where do these bands come from???

After PCR using Q5 pol, I ligate the insert into pCR-Blunt using T4 ligase. Transformation into DH10 is successful and a few colonies are mini-prepped. The plasmid is then cut with EcoR1-HF and...

31 December 2017 6,706 9 View

If a protein is easily aggregated during dialysis in HBS, does that infer that the protein would easily crystallize?

09 October 2016 3,548 7 View

Why are there no bands in my IP lanes?

07 August 2016 9,354 7 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Benefits of the 24 welled plate?

We have always used the 96 welled plate, the 48 welled plate but apparently there is much evidence on the 24 welled plate and above all its benefits.It states it is the same you would use on ELISA...

28 February 2021 1,149 1 View

Difficulty in cloning of 1kb gene into vector?

I am facing difficulties in cloning a 1kb gene into a vector (pJIT163). I have my gene on interest (GOI) in pUC57 and want to clone in pJIT163 using SalI and BamHI restriction sites. I am getting...

25 February 2021 3,221 7 View

Is RD-ES grow attached or in suspension or both?

We have a old RD-ES cell line which is grow attached, but we got another vial of RD-ES which is very easy to get detached and grow in sphere in the medium. Do you know which morphology is normal...

24 February 2021 5,183 1 View

What does it require to be ready for e-Commerce in developing countries?

Most developing countries are not ready for e-Commerce? What dimensions and indicators are there to measure e-commerce readiness of a nation? Please share framework or model and evaluation...

22 February 2021 8,132 6 View

Why are Sanger Sequencing reads interrupted in some edited clones?

I have a Crispr/Cas9 edited bulk population and I can briefly check the indel ratios in the bulk population using Sanger sequencing. After cloning single cells, I wanted to check the indel types...

18 February 2021 8,198 6 View

Hi, can anybody help me with the suggestions how can I amplify a stress induced gene of 1107 bp?

I am trying to amplify the TGA1 (Transcription factor 1 ) of Arabidopsis ,which is 1107 bp. I have treated the plants with Salycylic solution over night and extracted the RNA and make cDNA. I...

17 February 2021 7,802 2 View

Alternatives to traditional cloning of insert into backbone (dsDNA)?

Hi, I am stuck with a cloning experiment: The insert & backbone(bb) share one compatible end at 3' end(NotI) and one incompatible end at 5' end (XbaI for insert; HindIII for backbone). So i...

16 February 2021 1,664 3 View