miniaturisation of reverse transcription reaction volumes

Gene expression analysis can be performed using highly sensitive RNA sequencing methods, enabling very low sample inputs, even down to the single cell level. The use of mosquito liquid handlers provides reduced cost through miniaturisation of sample preparation and other benefits including:

  • low-volume sample prep, reduces reagent cost and sample input (gDNA/DNA/RNA)
  • positive displacement pipetting technology, essential in accurate and precise pipetting of small volumes and resulting in robust data
  • disposable tips eliminate cross-contamination
  • easy-to-use software provides fast reaction setup for low- to high-throughput applications
  • capable of full integration with other liquid handlers and ancillary equipment

case study: CHORI – low-cost SMARTer® ultra-low RNA input sample prep

The SMARTer® ultra® kit (Clontech, Mountain View, USA) is a cDNA synthesis kit specifically formulated for single-cell or ultra-low-input RNA-seq cDNA synthesis. For cost-saving purposes, in recent years, it has been of interest to sort cells using a cell sorter and then perform low volume cDNA synthesis, instead of costly microfulidics solutions. In order to reduce the cost and required sample input, this study set out to reduce the reaction volumes for the cell lysis, reverse transcription and cDNA synthesis using mosquito HTS liquid handler and compare the results with manual reaction set ups.

In order to reduce the cost and required sample input, mosquito HTS liquid handler was used to miniaturise the cDNA synthesis reaction volumes down to 12.5 and 6.25 μL. The study was carried out at Dr. Dario Boffelli  lab (Children Hospital of Oakland Research Institute, Oakland, CA).  The samples consisted of minimally diluted K562 single cells. As a comparison, a 25 μL reaction volume was set up manually with 10 of K562 cells. The results clearly show that lowering the reaction volumes by 2X, 4X and 8X (manually) did not affect the quality of cDNA synthesis, while providing 8 times cost saving per sample prep compared to the original sample prep volume.

Customer_spotlight_CHORI_table1

>> View the poster on methods for reduced cost and lower sample prep volumes for genetic analysis applications 

>> Get more information about mosquito HTS (25 nL – 1.2 µL)

>> Get more information about mosquito HV (0.5 – 5 µL) 

>> Get more information about mosquito X1 (hit picking)

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