Revision as of 20:32, 2 November 2016 by >Mzcai
CA12k_Oct2016 Probe Preparation
- Received CA12k_Oct2016_Oligos2Order_FINAL in 80ul TE Buffer
- 47.35ng/ul (3.79ug total)
- Calculate MW of 195 nt probe = 195nt * 303.7D/nt + 79D = 59,300.5 g/mol
- 47.35 ng/ul * nmol/59300.5ng = 0.0007985 nmol/ul = 0.8 pmol/ul = 0.8 uM
- Make 12.5ul aliquots in PCR strip tubes for future
Probeset
|
Amplification
|
# Probes
|
# Barcodes
|
# Genes
|
Human Brain
|
V4
|
4,978
|
391
|
368
|
Mouse Brain
|
V7
|
3,602
|
342
|
319
|
Mouse Embryo
|
V6
|
2,469
|
251
|
220
|
Lung Cancer
|
V8
|
105
|
105
|
85
|
Expansion PCR Test V4
Component
|
Volume
|
Seed oligo |
0
|
F/R V4 Primer Mix (10uM) |
2
|
2X Kapa MM |
12.5
|
H2O |
10.5
|
Total |
25
|
Component
|
Volume
|
TB12k_Apr2016 Seed oligo (100X Dilution) |
2.5
|
F/R V4 Primer Mix (10uM) |
2
|
2X Kapa MM |
12.5
|
H2O |
8
|
Total |
25
|
Component
|
Volume
|
Seed oligo (800nM) |
1.5
|
F/R V4 Primer Mix (10uM) |
2
|
2X Kapa MM |
12.5
|
H2O |
9
|
Total |
25
|
Component
|
Volume
|
Seed oligo (800nM) |
3.125
|
F/R V4 Primer Mix (10uM) |
2
|
2X Kapa MM |
12.5
|
H2O |
7.375
|
Total |
25
|
Program
95C 30sec -> (95C 5sec -> 55C 30sec-> 72C 45sec) x 30 -> 72C 2min -> 15C hold
Expansion PCR V4