Bead Testing on Old and New Beads

Date Performed: May 6th, 2021

Samples selected for testing: 448-2, 438, 779, 877
Samples from next round of sequencing

• Thawed, vortexed, spun down
• Made two sets of tubes following chart below
• one is for testing the new beads the other is for testing old beads\

Qubit before clean (after dilution above)

• -N denotes “new” beads, -O denotes “old” beads

Sample	Run 1	Run 2
S1	196.05
S2	22448.42
448-N	26.4	26.4
438-N	15.9	15.4
779-N	15.3	15.2
877-N	29.2	28.0
448-O	26.6	26.2
438-O	14.2	14.2
779-O	14.2	14.1
877-O	26.0	26.8

Followed 1.8X bead clean, only change is cleaned with 150ul ethanol instead of 200ul
Eluted in 20ul water because I removed 1ul per sample for Qubit

Post Bead Clean Qubit

Sample	Run 1	Run 2
S1	172.46
S2	22546.19
448-N	23.8	23.6
438-N	11.8	11.8
779-N	12.7	12.6
877-N	18.4	18.1
448-O	20.2	20.2
438-O	9.10	8.98
779-O	9.32	9.28
877-O	14.8	14.5

These results didn’t look great. Looked like a lot was lost.
Next I tested three samples by diluting, quantifying, bead cleaning (1.8X), then running in thermocycler.
Added 2ul cutsmart buffer before putting in thermocylcer

Sample	Volume for 400ng DNA	Volume up to 21ul Water
413	4.0	17.0
440	4.0	17.0
768	4.0	17.0

Pre-Thermocylcer Assay

Sample	Run 1	Run 2
S1	202.64
S2	24478.58
413-O	17.8	17.2
440-O	20.2	19.8
768-O	13.5	10.6
413-N	14.9	14.8
440-N	20.6	20.2
768-N	11.0	16.4

All Results Below (Some duplicate info)