Target Knock Down and phenotypic changes in the test system following mRNA knock down
I would like to know the level of expression of my target in a cell line

RNAx will test the expression of the target in a cell line of your choice

This is a typical set up of such an experiment:

  • The customer delivers the accession number of the target to RNAx
The experimental set-up consists of the following steps:
  • Design of oligonucleotides for real time PCR
  • RNA isolation
  • Performance of real time PCR
  • Compilation of data
  • Delivery of the data to the customer

...back

 
 
Target Knock Down and phenotypic changes in the test system following mRNA knock down
I would like to inhibit the expression of a target using siRNAs. Quantification should be done on the mRNA level

RNAx will design siRNA oligos and perform knock down experiments. The measurement of the knock down will be done by real time PCR

Hier ein Beispiel für einen typischen Verlauf eines Knock Down ExperimentsThis is a typical set up of such an experiment:

  • The customer delivers to RNAx the siRNA and information on the expression of the target sequence in cell lines
  • RNAx performs the knock down experiments in established in-house cell lines, if the target sequence is expressed in these cell lines
  • If the target sequence is not expressed in in-house cell lines, RNAx performs an adaptation of the customer's cell line to the automated experimental set up
The experimental set-up consists of the following steps:
  • Test of expression of the target sequence in two in-house cell lines
  • Design of oligonucleotides for real time PCR
  • Design of two siRNA oligos per target
  • Transfection of the customer's siRNA in one chosen cell line in triplicates (three experiments)
  • In parallel, one control experiment will be carried out
  • RNA isolation
  • Real time PCR
  • Compilation of data
  • Delivery of the data to the customer

...back

 
 
Target Knock Down and phenotypic changes in the test system following mRNA knock down
I would like to inhibit the expression of a target using siRNAs. Knock down should be shown on the protein level

RNAx will design siRNA oligos and perform knock down experiments. The measurement of the knock down will be done by real time PCR and Immunoblot.

This is a typical set up of such an experiment:

  • The customer delivers to RNAx the siRNA and information on the expression of the target sequence in cell lines
  • RNAx performs the knock down experiments in established in-house cell lines, if the target sequence is expressed in these cell lines
  • If the target sequence is not expressed in in-house cell lines, RNAx performs an adaptation of the customer's cell line to the automated experimental set up
The experimental set-up consists of the following steps:
  • Test of expression of the target sequence in two in-house cell lines
  • Design of oligonucleotides for real time PCR
  • Design of two siRNA oligos per target
  • Transfection of the customer's siRNA in one chosen cell line in triplicates (three experiments)
  • In parallel, one control experiment will be carried out
  • RNA isolation
  • Real time PCR
  • Performance of an Immunoblot to show protein knock down
  • Compilation of data
  • Delivery of the data to the customer

...back

 
 
Target Knock Down and phenotypic changes in the test system following mRNA knock down
I would like to inhibit the expression of a target using siRNAs. Phenotypic changes should be shown using a functional assay

RNAx will design siRNA oligos and perform knock down experiments. The measurement of the knock down will be done by real time PCR and functional assays.

This is a typical set up of such an experiment:

  • The customer delivers to RNAx the siRNA and information on the expression
  • RNAx performs the knock down experiments in established in-house cell lines, if the target sequence is expressed in these cell lines
  • If the target sequence is not expressed in in-house cell lines, RNAx performs an adaptation of the customer's cell line to the automated experimental set up
The experimental set-up consists of the following steps:
  • Test of expression of the target sequence in two in-house cell lines
  • Design of oligonucleotides for real time PCR
  • Design of two siRNA oligos per target
  • Transfection of the customer's siRNA in one chosen cell line in triplicates (three experiments)
  • In parallel, one control experiment will be carried out
  • RNA isolation
  • Real time PCR
  • Performance of the functional assay(s) (For a list of avalilable assays klick:...here)
  • Compilation of data
  • Delivery of the data to the customer

...back

 
Functional Validation of shRNAs and Target Knock Down
I would like to identify a functioning shRNA and perform a target knock down

RNAx will design shRNA-encoding DNA-oligonucleotide pairs and perform knock down experiments. The measurement of the knock down will be done by real time PCR.

This is a typical set up of such an experiment:

  • The customer delivers to RNAx the accession number of the target
  • RNAx performs the knock down experiments in established in-house cell lines, if the target sequence is expressed in these cell lines
  • If the target sequence is not expressed in in-house cell lines, RNAx performs an adaptation of the customer's cell line to the automated experimental set up
The experimental set-up consists of the following steps:
  • Test of expression of the target sequence in in-house cell lines
  • Design of oligonucleotides for real time PCR
  • Design of four shRNA-encoding DNA-oligonucleotide pairs per target
  • Cloning of shRNA-encoding DNA-oligonucleotide pairs into vector and verifying constructs by sequencing
  • Generation of Virus by transfection of the shRNA constructs together with packaging plasmids (triplicates, three times independently, respectively)
  • In parallel, one control experiment will be carried out
  • Infection of one chosen cell line with virus
  • RNA isolation
  • Real time PCR
  • Compilation of data
  • Delivery of the data to the customer

...back

RNAx offers
RNAi-Screens for Target Identification with Subsequent Hit Validation

RNAx offers to run automated RNAi-screens. This way, you can take advantage of this technology, even if you do not have the expertise or equipment to perform such screens.

Here you can see a short description of such a screen which, of course, will completely be customized to meet your research needs ...read more

Target Knock Down and phenotypic changes in the test system following mRNA knock down
I would like to know the level of expression of my target in a cell line ...read more

I would like to inhibit the expression of a target using siRNAs. Quantification should be done on the mRNA level ...read more

I would like to inhibit the expression of a target using siRNAs. Quantification should be done on the protein level ...read more

I would like to inhibit the expression of a target using siRNAs. Quantification should be done using a functional assay ...read more
Functional Validation of shRNAs and Target Knock Down
I would like to identify a functioning shRNA and perform a target knock down ...read more