A method for specifically and efficiently quantifying the expression of targeted RNA variants with specific terminal sequences suitable to identify multiple isoforms bearing complex heterogeneity in terminal sequences by hybridizing a 5′-Dbs-adapter to the 5′-end of target RNAs, wherein the 5′-Dbs-adapter has a stem-loop structure whose protruding 5′-end base-pairs with the 5′-end of target RNAs, and wherein the loop region of 5′-Dbs-adapter contains a base-lacking spacer which will terminate reverse transcription in a subsequent step; hybridizing a 3′Db-adapter to the 3′-end of target RNAs, wherein the 3′-Db-adapter has a stem-loop structure whose protruding 3′-end base-pairs with the 3′-end of target RNAs; ligating both adapters with target RNAs by RN12 ligation to form a “dumbbell-like” structure; and, amplifying and quantifying the ligation product by RT-PCR.

A method for specifically and efficiently quantifying the expression of targeted RNA variants with specific terminal sequences suitable to identify multiple isoforms bearing complex heterogeneity in terminal sequences by hybridizing a 5′-Dbs-adapter to the 5′-end of target RNAs, wherein the 5′-Dbs-adapter has a stem-loop structure whose protruding 5′-end base-pairs with the 5′-end of target RNAs, and wherein the loop region of 5′-Dbs-adapter contains a base-lacking spacer which will terminate reverse transcription in a subsequent step; hybridizing a 3′Db-adapter to the 3′-end of target RNAs, wherein the 3′-Db-adapter has a stem-loop structure whose protruding 3′-end base-pairs with the 3′-end of target RNAs; ligating both adapters with target RNAs by RN12 ligation to form a “dumbbell-like” structure; and, amplifying and quantifying the ligation product by RT-PCR.

Technology provided herein relates in part to methods, processes and apparatuses for non-invasive assessment of genetic variations.

Technology provided herein relates in part to methods, processes and apparatuses for non-invasive assessment of genetic variations.

The present disclosure provides a novel approach, recomSELEX, that highly integrate mutually supportive recombination and computational methods for aptamer selection and identification. The recomSELEX approach comprises a recombinatorial SELEX platform for aptamer screening that exponentially increases the sequence space that is explored by incorporation of a DNA shuffling step that allows recombination between aptamers. Subsequently, the recombinatorial SELEX platform can also be employed to develop new and optimize already existing aptamers. The recomSELEX further comprises a computational SELEX platform with a constrained genetic algorithm (GA) to identify potential aptamers that are stable and have the desired specificity and affinity of a target.

The present disclosure provides a novel approach, recomSELEX, that highly integrate mutually supportive recombination and computational methods for aptamer selection and identification. The recomSELEX approach comprises a recombinatorial SELEX platform for aptamer screening that exponentially increases the sequence space that is explored by incorporation of a DNA shuffling step that allows recombination between aptamers. Subsequently, the recombinatorial SELEX platform can also be employed to develop new and optimize already existing aptamers. The recomSELEX further comprises a computational SELEX platform with a constrained genetic algorithm (GA) to identify potential aptamers that are stable and have the desired specificity and affinity of a target.

The present disclosure provides a novel approach, recomSELEX, that highly integrate mutually supportive recombination and computational methods for aptamer selection and identification. The recomSELEX approach comprises a recombinatorial SELEX platform for aptamer screening that exponentially increases the sequence space that is explored by incorporation of a DNA shuffling step that allows recombination between aptamers. Subsequently, the recombinatorial SELEX platform can also be employed to develop new and optimize already existing aptamers. The recomSELEX further comprises a computational SELEX platform with a constrained genetic algorithm (GA) to identify potential aptamers that are stable and have the desired specificity and affinity of a target.

The invention provides methods for simultaneously amplifying multiple nucleic acid regions of interest in one reaction volume as well as methods for selecting a library of primers for use in such amplification methods. The invention also provides library of primers with desirable characteristics, such as minimal formation of amplified primer dimers or other non-target amplicons.

The invention provides methods for simultaneously amplifying multiple nucleic acid regions of interest in one reaction volume as well as methods for selecting a library of primers for use in such amplification methods. The invention also provides library of primers with desirable characteristics, such as minimal formation of amplified primer dimers or other non-target amplicons.

The invention relates to a method for detecting residual, undifferentiated pluripotent stem cells (PSCs) in a culture of cells differentiated from PSCs, the method comprising: culturing the cells on a substrate coated with laminin-521 and E-cadherin in a medium comprising a ROCK inhibitor; quantitating in the cultured cells expression of a marker of residual, undifferentiated PSCs; and comparing the marker expression in the cultured cells with the marker expression in a reference culture of cells comprising a known proportion of PSCs, wherein lower marker expression in the culture of cells than marker expression in the reference culture of cells indicates absence of residual, undifferentiated PSCs in the cultured cells or presence of residual, undifferentiated PSCs in the cultured cells at a proportion lower than the known proportion of PSCs in the reference culture of cells. The invention also relates to a method for manufacturing a therapeutic composition and a method for treating or preventing a condition in a subject.