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

More Shanice Chiang's questions See All

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 9,345 4 View

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 8,610 9 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 8,764 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 6,103 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 3,617 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 5,368 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 5,478 4 View

How does one extract inclusion bodies from E. coli?

what is the most effective method here

07 August 2014 8,345 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 3,715 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 1,768 4 View

How to compare two groups with only two measurements?

Hiiiii everyone! I have an enquiry on statistical analysis. I was looking for many forum and it's still cannot solve my problem. I want to compare means of two groups of data but only with two...

03 March 2021 8,796 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

What kind of fluid could I use for a pitfall trap that also does not invalidate molecular testing?

Hi, I want to start testing pitfall trap to obtain ants samples, but I need to conduct molecular analysis on those insects. So, what kind of fluid can I use? Ethanol expires too early and I need...

03 March 2021 5,978 5 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

What's the best way to measure growth rates in House sparrow chicks from day 2 to day 10?

What's the best way to measure growth rates in House sparrow chicks from day 2 to day 10? Since, the growth curve from day 2 to 10 won't be like the "Logistic curve" it might not follow logistic...

03 March 2021 1,401 3 View

Do we need to write protocol for a second study of a systematic review that has been registered and published?

I have conducted and published a systematic review and meta-analysis research with the topic related to public health and health pomotion (protocol was registed in PROSPERO). Now we want to...

03 March 2021 8,920 3 View

How to increase a model accuracy ?

dear community, my model is based feature extraction from non stationary signals using discrete Wavelet Transform and then using statistical features then machine learning classifiers in order to...

03 March 2021 6,994 5 View

Do I need to do linear regression before running mediation analysis using PROCESS MACRO by Hayes ?

I just wanted to check if I need to run a linear regression separately if I am using PROCESS MACRO to run mediation analysis. Thank you.

02 March 2021 4,359 3 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Is There Any Feasible Method To Test Fluorescent Compounds Other Than Fluorescence Spectrometers ??

Is There Any Feasible Method To Test The Efficiency Of Fluorescent Compounds Other Than UV Spectrometers ? Suggestions Would Be Appreciated !

02 March 2021 5,785 3 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