Can a tet-off sysytem be replaced by a tet-on one?

07 July 2014 1 8K Report

I have a plasmid in which the TetO2 operator sequence is present in the promoter for the gene. The manual says that it is supposed to be used for the tet-off system. Is the binding sequence for the tetR in the tet-off and tet-on systems same? Can the above plasmid be used for a tet-on system? If someone has any relevant study material on this, please do share.

Sanath Kumar Janaka

The tet-off and tet-on systems are different in that you will need to use doxycycline to allow expression of your gene of interest in a tet-on system. Otherwise, the operator sequence is the same for both. You could switch them back and forth.

Badges
Science topic

More Debosree Pal's questions See All

My antibody is not pulling down the protein of interest during immunoprecipitation. What should I do?

I have an in-house raised antibody against my protein of interest which I have purified from the serum by the caprylic acid method and further confirmed on a sds gel. I wanted to use it for...

11 December 2015 2,638 7 View

Has anyone worked on embryonic germ cells as a model system?

I want to work on embryonic germ cells. Can anyone suggest from where I can procure these cells? Also, does anyone know the difference between embryonic germ cells and primordial germ cells

01 February 2015 1,301 1 View

Can anyone share with me a protocol for the differentiation of mouse embryonic stem cells into primordial germ cells?

Hi, I am looking for a protocol to differentiate mouse embryonic stem cells into primordial germ cells. Can anyone provide me with a convenient protocol? How long can the differentiated cells be...

09 October 2014 9,618 5 View

Can R1 embryonic stem cells be grown in 2i+LIF medium?

Hello, I am growing R1-ESCs in high glucose DMEM, 15%FBS, sodium pyruvate, non-essential amino acids and L-glutamine. Can these cells be grown in 2i+LIF medium? How many passages do I need to grow...

09 October 2014 7,158 4 View

Does anyone have experience with an inducible lentivirus based shRNA expression system for mouse embryonic stem cells?

I am planning to perform shRNA -mediated knockdown of my gene of interest using an inducible lentiviral based system. I am working on mouse embryonic stem cells. Can someone suggest a suitable...

07 August 2014 5,959 7 View

Can anyone suggest a suitable tet-on inducible lentiviral plasmid for shRNA expression?

I want to express shRNA against my gene of interest in a tetracycline regulatable manner (tet-on system). Can anyone suggest some suitable plasmids for the same? I have shRNAs with 5'BglII and...

06 July 2014 3,461 3 View

Does the method of homogenization of tissues bring about a change in downstream PCR results?

I was trying to see the expression of a gene in post-implantation mouse embryos from 9.5-13.5dpc. The preferred technique is obviously whole embryo in-situ hybridization but, I first thought of...

04 May 2014 2,166 4 View

Can anyone suggest how to store hybridization buffer?

I am using a hybridization buffer for in-situ hybridization which has the following components: 50% formamide, 5X SSC, yeast RNA, heparin and 1% SDS. Storing the buffer at 4 degrees or -20 degrees...

04 May 2014 3,316 6 View

How tricky is it to isolate and culture primary mouse embryonic fibroblasts?

I have been trying to culture mouse embryonic fibroblasts for quite a while now but have been facing a lot of problems with the same. I isolated the fibroblasts from E13.5 old mouse embryos (CD-1...

03 April 2014 7,618 7 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

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 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

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

Irradiation data for JAPAN?

Hi, Could you help me to get the latest (2019, 2020) solar resource data for a particular site in JAPAN? PV GIS and NSRDB data are not in reach to JAPAN, I'm looking for some other database for...

03 March 2021 961 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