Abstract: A method for determining one or more tumour properties in a subject with a tumour, the method comprising: —seeding a cell-free scaffold obtained from the tumour with cancer cells; —culturing the cancer cells in the scaffold; —assaying the cultured cancer cells for the presence of target molecules indicative of the expression of one or more genes in the cells; and —determining one or more tumour properties based on the results of the assay.

Abstract: Cell-free scaffolds derived from tumors in patients are used as in vitro cancer models and also provide information of the tumor, from which it is derived, including its susceptibility to cancer treatment. The cell-free scaffolds are also used as a predictive tool in assessing cancer treatment efficacy and in identifying immunotargets and biomarkers for cancer therapy.

Abstract: A method for determining one or more tumour properties in a subject with a tumour, the method comprising:—seeding a cell-free scaffold obtained from the tumour with cancer cells;—culturing the cancer cells in the scaffold;—assaying the cultured cancer cells for the presence of target molecules indicative of the expression of one or more genes in the cells; and—determining one or more tumour properties based on the results of the assay.

Abstract: The present invention determines lymphocyte clonality by contacting a sample comprising nucleic acid molecules (1) of lymphocytes with forward and reverse primers (10, 20) and amplifying the nucleic acid molecules (1) by performing PCR pre-amplification to form barcoded PCR products (50). The barcoded PCR products (50) are amplified using adapter-specific forward and reverse primers (30, 40) in a PCR application into amplified barcoded PCR products (60), which are sequenced. The sequence reads are demultiplexed, mapped to respective TCR or BCR clonotypes and used to determine lymphocyte clonality for the sample. The forward and/or reverse primers (10, 20) are barcoded by comprising UMIs (14, 24) protected inside hairpin loops.

Abstract: A method for determining one or more tumour properties in a subject with a tumour, the method comprising:—seeding a cell-free scaffold obtained from the tumour with cancer cells;—culturing the cancer cells in the scaffold;—assaying the cultured cancer cells for the presence of target molecules indicative of the expression of one or more genes in the cells; and—determining one or more tumour properties based on the results of the assay.

Abstract: The present invention is directed to a method for performing RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M Guanidine Thiocyanate, b) diluting the sample to an extend such that Guanidine Thiocyanate is present in a concentration of about 30 to 50 mM, e) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of thermocycling protocol and monitoring amplification of the first strand cDNA in real time, characterized in that steps a) to c) are done in the same vessel.

Abstract: The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M Guanidine Thiocyanate, b) diluting the sample to an extent such that Guanidine Thiocyanate is present in a concentration of about 30 to 50 mM, c) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of a thermocycling protocol and monitoring amplification of the first strand cDNA in real time, characterized in that steps a) to c) are done in the same vessel.

Abstract: The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M guanidine thiocyanate, b) diluting the sample to an extend such that guanidine thiocyanate is present in a concentration of about 30 to 50 mM, c) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of a thermocycling protocol and monitoring amplification of the first strand cDNA in real time.

Abstract: The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M Guanidine Thiocyanate, b) diluting the sample to an extend such that Guanidine Thiocyanate is present in a concentration of about 30 to 50 mM, c) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of a thermocycling protocol and monitoring amplification of the first strand cDNA in real time, characterized in that steps a) to c) are done in the same vessel.

Abstract: The present invention is directed to a method for performing an RT-PCR for amplifying a target RNA comprising the steps of a) lysis of a cellular sample which is supposed to contain the target RNA with a lysis buffer comprising between 0.2 M and 1 M guanidine thiocyanate, b) diluting the sample to an extend such that guanidine thiocyanate is present in a concentration of about 30 to 50 mM, c) reverse transcribing in the presence of a mixture of first strand cDNA synthesis primers, the mixture consisting of oligo dT primers and random primers, and d) subjecting the sample to multiple cycles of a thermocycling protocol and monitoring amplification of the first strand cDNA in real time.