Continued with templated bead prep from ePCRs for these libraries. Samples were processed according to the ABI “Templated Bead Preparation Guide” following the “full-scale” protocol.
To see explanations of the various calculations below, see the “SOLiD Bead Titration – Herring fragmented cDNA library 3LHSITK09 (CONTINUED from ePCR yesterday)” from 20100107.
2LHKOD09:
Initial counts: 109, 129, 112, 115 —- Avg. = 116.25 beads/square
Beads: 116.25 x 10 x 25 x 200 = 5.8125×10^6 beads/uL x 200uL = 1.1625×10^9 beads
Templated beads counts: 205, 199, 197, 210 —– Avg. = 210 beads/square
Templated beads: 210 x 10 x 25 x 10 = 5.06875×10^5 beads/uL x 400uL = 2.0275×10^8 beads
Efficiency: 17.44%
4LHTOG09:
Initial counts: 124, 124, 117, 104 —– Avg. = 117.25 beads/sqaure
Beads: 117.25 x 10 x 25 x 200 = 5.8625×10^6 beads/uL x 200uL = 1.1725×10^9 beads
Templated beads counts: 139, 135, 145, 140 —- Avg. = 139.75 beads/square
Templated beads: 139.75 x 10 x 25 x 10 = 3.49375×10^5 beads/uL x 400uL = 1.3975×10^8 beads
Efficiency: 11.92%
6LHPWS09:
Initial counts: 135, 106, 123, 124 —- Avg. = 122 beads/square
Beads: 122 x 10 x 25 x 200 = 6.1×10^6 beads/uL x 200uL = 1.22×10^9 beads
Templated beads counts: 141, 171, 164, 170 —– Avg. = 161.5 beads/square
Templated beads: 161.5 x 10 x 25 x 10 = 4.0375×10^5 beads/uL x 400uL = 1.615×10^8 beads
Efficiency: 13.24%
All beads were stored @ 4C until ready for bead deposition and work flow analysis run.
Results:
Rhonda Morales (from Ginger’s lab who is responsible for running/maintaining the SOLiD at the CEG) says the numbers on all samples look perfect! Will proceed to work flow analysis once Jesse’s samples are ready (ETA of Jan. 27th, 2010).
Here are reagent lot numbers for the Bead Titration.