CirclSeq
Automatable and reliable preparation of a DNA library in a one-vial reaction for unbiased quantification of mRNA
One-vial reaction
Enables the unbiased amplification-free gene expression analysis from cells or biological samples
Reflects the real status of the cell
Pooling of samples is possible after the first reaction step
* The technique is
* automatable
* reliable
* time-saving
* cost-saving and
* amenable to high throughput
This method offers the possibility of an automatable, time- and cost-saving high throughput sequencing technique of mRNA or other nucleic acid molecules without the need for prior amplification.
The blocking of all 3' ends by the untemplated incorporation of di-desoxy-TTP (ddTTP) by terminal desoxynucleotidyl transferase hinders non-elongated primers from circularization and thus from later sequencing. Additional purification steps to remove the primers are not necessary anymore. cDNA digestion at dUTP sites still gives rise to 3'OH ends that can be circularized and analyzed by deep sequencing subsequently.
Thus, this novel method allows much higher throughput and maximizes the overall yield and the fraction of sequencing reads corresponding to real mRNA templates.
Licensing
30/10/2015 00:00:00
US201514929079 20151030
- international:
C12N15/10; C12Q1/68
- cooperative:
default
C12N15/1065; C12Q1/6806
C-sets
C12Q1/6806, C12Q2521/107, C12Q2521/301, C12Q2525/119, C12Q2525/307, C12Q2527/137
Patent application
4031
德国
