ePCR was performed following ABI’s “full scale” protocol, using 1pM of SOLiD cDNA library.
Templated bead preparation was performed according to the “full scale” protocol.
Bead counts are calculated as follows:
Avg bead count x # hemacytometer squares x volume in hemacytometer (uL) x dilution factor = beads/uL x suspension volume (uL) = total beads
Initial Bead count: (1:200 dilution)
Lean: 126, 138, 122, 138 Avg. = 131
Lean: 131 x 25 x 10 x 200 = 6.55x10^6 beads/uL x 200uL = 1.31x10^9 beads
Templated Bead counts (1:10 dilution)
Lean: 165, 171, 186, 160 Avg. = 170.5
170.5 x 25 x 10 x 10 = 426250 beads/uL x 400uL = 1.705x10^8 beads
Percent Recovery Templated Beads
Lean: (1.705x10^8 beads)/(1.31x10^9 beads) x 100 = 13.02% recovery
Results: Yield is significantly higher than the previous preparation performed with this sample. The percent recovery falls into the desired range of 5-15%, so things look good there, too. Will contact Rhonda and get info regarding when this and the other 7 samples can go on a run.