Question: How much of my cDNA is used for library prep on the Chromium Connect?
3' Gene Expression
- When the mid-run QC is selected, the Chromium Connect will dispense 25μl aliquots of cDNA into a strip tube for the user to perform subsequent QC steps on a Bioanalyzer or Tapestation off-deck. A 15μl fraction (~40%) of the cDNA remains on deck for fragmentation and library preparation.
When the mid-run QC is NOT selected, the cDNA will remain on deck and will not be available for subsequent QC steps. A 15μl aliquot (~40%) of the cDNA will be still be used for fragmentation and library preparation.
Note the volume is not adjustable if low cDNA yield is observed. However, the SI-PCR cycles can be adjusted during mid-run QC.
5' Gene Expression
- Chromium Connect dispenses 15μl aliquots of cDNA into a strip tube for the user to perform subsequent QC steps on a Bioanalyzer or Tapestation off-deck. The volume of cDNA (5μl-22μl) used for fragmentation and library construction will vary depending on the amount needed to target 60ng input. If the volume for 60ng exceeds 22μl, carry forward ONLY 22μl sample into library construction and adjust the SI PCR cycles accordingly.
- If selecting the cDNA only feature, the entire 40ul of cDNA will be transferred into the strip tube. 2ul of cDNA will be used for V(D)J amplification.
Products: Chromium Connect