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 = 5.8125x10^6 beads/uL x 200uL = 1.1625x10^9 beads
Templated beads counts: 205, 199, 197, 210 —– Avg. = 210 beads/square
Templated beads: 210 x 10 x 25 x 10 = 5.06875x10^5 beads/uL x 400uL = 2.0275x10^8 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 = 5.8625x10^6 beads/uL x 200uL = 1.1725x10^9 beads
Templated beads counts: 139, 135, 145, 140 —- Avg. = 139.75 beads/square
Templated beads: 139.75 x 10 x 25 x 10 = 3.49375x10^5 beads/uL x 400uL = 1.3975x10^8 beads
Efficiency: 11.92%
6LHPWS09:
Initial counts: 135, 106, 123, 124 —- Avg. = 122 beads/square
Beads: 122 x 10 x 25 x 200 = 6.1x10^6 beads/uL x 200uL = 1.22x10^9 beads
Templated beads counts: 141, 171, 164, 170 —– Avg. = 161.5 beads/square
Templated beads: 161.5 x 10 x 25 x 10 = 4.0375x10^5 beads/uL x 400uL = 1.615x10^8 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).
