How can I check if the cloned gene is in the frame?

More Shanice Chiang's questions See All

Would I be able to see a difference between a induced and uninduced empty pET vector?

Hi people, I am having a lot of difficulties in my protein expression. I am wondering if my induction has failed to work? Too bad, i dont have any positive control. So i am using the empty...

09 October 2014 8,665 9 View

Is it possible to replace yeast nitrogen base with other materials?

I am going to do induction in Pichia pastoris. But my lab does not have yeast nitrogen base. Just wondering if I can substitute with some other things.

09 October 2014 9,470 4 View

Gene with high GC content is difficult to be expressed in E. coli. How do you define high here?

People I need some help here.

09 October 2014 8,822 0 View

how to tell if protein induction works?

I am not sure if my induction have worked. Is it possible to do a RNA miniprep. IF RNA successful extracted- means induction has worked.

09 October 2014 6,179 0 View

Can anyone recommend a website that can predict RNase E cutting site?

Do you know of any website which would predict RNAse E cutting site? or how to predict it? We are having problems expressing a foreign mRNA in E.coli and one possibility was that the mRNA was...

09 October 2014 3,671 3 View

D-lactose monohydrate versus alpha lactose monohydrate for use in autoinduction media

I need lactose for my autoinduction media for E.coli. Just wondering if I could use the D-lactose monohydrate because I can't find any alpha-lactose monohydrate in my lab. Are they the same?

08 September 2014 5,424 8 View

Anyone have experience in autoinduction?

I have cloned lipase gene into pET28+ and express it in Rosetta(DE3)pLyss. I have tried to induce with IPTG but I can't spot any difference between the clone and the empty pET. I varied the...

08 September 2014 5,549 4 View

How does one extract inclusion bodies from E. coli?

what is the most effective method here

07 August 2014 8,404 2 View

Can anyone help with induction in E. coli?

Dramatic induction was observed, but no desired protein band was observed. I run an empty pET as a control. There doesnt seem to be difference with the empty pET and pET with insert. I have no...

07 August 2014 3,782 13 View

How do you choose a good antibody for western blot?

I have cloned my lipase gene from rice in pET-28(+) His-tag vector. Was wondering how do I choose/design the primary and secondary antibodies for western blot. I am totally new to this. Was hoping...

06 July 2014 1,835 4 View

How to learn more about SPSS and its Application?

I would like to learn more about SPSS and Its application especially in regards to data analysis. Please suggest me how I can learn more about it. Thank you so much.

11 August 2024 9,101 4 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

Baseline drift in HPLC? What causes this?

Hello, Why do i see this baseline drift when i compare my blank (black) to the sample (blue)? Any suggestions as to why this happened? Thank you!

11 August 2024 3,770 4 View

Text-Communication from the M1 Hand Area using BCI—and then there is Elon Musk?

Willett, Shenoy et al. (2021) have developed a brain computer interface (BCI) that used neural signal collected from the hand area of the motor cortex (area M1) of a paralyzed patient. The...

10 August 2024 7,180 0 View

Has anyone applied Python in the field of textile engineering for data analysis, automation, or smart textiles?

I'm currently exploring the application of Python in textile engineering, specifically in areas like data analysis, process automation, and the development of smart textiles. I'm interested in...

10 August 2024 7,429 2 View

Is there an English Translation of the Carl Moller text: ZUR VERGLEICHENDEN ANATOMIE DER SILURIDEN?

I recently came across an anatomy text by Carl Moller that was published in 1915 but it is in German or Dutch neither of which I can understand. I would like to know if there is an English...

10 August 2024 4,347 1 View

How can I use the cif data obtained from rietveld refinement extracted via gsas2, for microstructural analysis using ETEX software?

09 August 2024 7,718 0 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

Does crude extraction using NaOH and Tris work well with Fungi?

I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you

08 August 2024 4,733 2 View

Marcin Nowicki

First you need to sequence the constructs, using the pET-specific primers. Then, yes, you do work with the resultant sequences to ensure in-frame status.

Shanice Chiang

Thanks DR. Nowicki

Pranali Deore

Try doing cloning in Nco-1 or nde-1 , these sites at 5' end gives ATG so that your construct is in frame.

Muhammad Owais Shahid

I have the same question, but these answers doesn't help me