Journal: Biomicrofluidics

Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform

doi: 10.1063/1.4991005

Figure Lengend Snippet: The Cell-SELEX process performed on a microfluidic chip. (a) Incubation and mixing of the single-stranded DNA (ssDNA) library with target cell-magnetic bead complexes in binding buffer and fetal bovine serum (FBS) in the positive selection chamber. (b) Washing and removal of unbound ssDNA using the washing buffer and removal of wastes through the waste collection chamber. (c) Thermal release of bound ssDNA and application of a magnetic field to isolate the cells and transfer the bound ssDNA to the negative selection chamber. (d) Negative selection in the negative selection chamber with negative control cell-magnetic bead complexes in binding buffer. (e) Magnetic separation of negative control cell-magnetic bead complexes, and transfer of supernatant to the PCR chamber. (f) PCR amplification of selected sequences. The amplified product was then transferred to the positive selection chamber for the next round of SELEX. Blue arrows indicate the micro-chamber in which the respective reaction step was carried out.

Article Snippet: Capillary electrophoresis (CE)-based SELEX, 18 sol-gel-based SELEX, 19 and magnetic bead-based SELEX 20–22 have all been integrated into microfluidic systems, resulting in the rapid and highly efficient isolation of aptamers.

Techniques: Incubation, Binding Assay, Selection, Negative Control, Amplification

Journal: Biomicrofluidics

Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform

doi: 10.1063/1.4991005

Figure Lengend Snippet: Agarose gel electrophoresis: (a) PCR products were electrophoresed on an agarose gel (2%) and stained with ethidium bromide. For panels (a) and (b), lane L = 50-bp DNA ladder, lane N = ddH2O (negative control), lane P = 1 μM of ssDNA library, and lanes S1 to S6= PCR products (30 cycles) obtained after 1 to 6 rounds of SELEX, respectively, with HuCCT-1 (a) and SNU-478 (b) cells. (c) Gel electrophoresis analysis after a second round of negative selection performed with MMNK-1 cells and WBCs. Lanes S and H depict PCR products after negative selection with SNU-478 and HuCCT-1 cells, respectively.

Article Snippet: Capillary electrophoresis (CE)-based SELEX, 18 sol-gel-based SELEX, 19 and magnetic bead-based SELEX 20–22 have all been integrated into microfluidic systems, resulting in the rapid and highly efficient isolation of aptamers.

Techniques: Agarose Gel Electrophoresis, Staining, Negative Control, Nucleic Acid Electrophoresis, Selection

Journal: Biomicrofluidics

Article Title: Automated selection of aptamers against cholangiocarcinoma cells on an integrated microfluidic platform

doi: 10.1063/1.4991005

Figure Lengend Snippet: Melting curves analysis of the ssDNA pools after each round of SELEX. (a) Melting curve of SELEX rounds 1 to 6 for SNU-478 cell line. (b) Melting curve of SELEX rounds 1 to 6 for HuCCT-1 cell lines. The RT-PCR products of the 5th and 6th cycles depicted a melting temperature of 84.88 °C and 85.02 °C.

Article Snippet: Capillary electrophoresis (CE)-based SELEX, 18 sol-gel-based SELEX, 19 and magnetic bead-based SELEX 20–22 have all been integrated into microfluidic systems, resulting in the rapid and highly efficient isolation of aptamers.

Techniques: Reverse Transcription Polymerase Chain Reaction