Entry title
- PCR - validation of Chromatin sensor insert
PCR - validation of Chromatin sensor insert
- Designed primers to amplify three overlapping regions across the transgenes: Chromosensor1, Chromosensor 2
- chrsn1 f1/ chrsn1 r1; Validation region 1; sensor 1 (and 2)
- chrsn1 f2/ chrsn1 r2; Validation region 1; sensor 1 (and 2)
- Round 1: Control templates to test primer function
- U-2 OS genomic DNA (neg)
- Chromosensor 1 plasmid DNA
- Chromosensor 2 plasmid DNA
- Round 2: Genomic DNA prepped from candidate clonal lines
- To be done after chosing a single primer pair per validation region
- BD###, Chromosensor 1
- BD###, Chromosensor 1
- BD###, Chromosensor 2
- BD###, Chromosensor 2
ROUND 1 PCR: Neg, pos controls
| Reagent
|
Vol
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Mix* (x #)
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Expected:
1. CMV = 588
|
Hover name
5 μL/lane; 1% agarose; Ladder
|
| genomic OR plasmid DNA |
1.0 / 0.5 |
7.0 / 4.5
|
| 10 μM forward primer |
1.0 |
---
|
| 10 μM reverse primer |
1.0 |
---
|
| 2x GoTaq green |
10.0 |
70.0
|
| dH2O |
7.0 / 7.5 |
49.0 / 52.5
|
| |
20.0 μL |
---
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Note: *Prep one Mix per template
- For each template, aliquot 18.0 master mix into 6 tubes
Thermal cycler: Labnet - GOTAQ
- 95°C 3 min.
- 30x[95°C, 30 sec; 57°C 30 sec; 72°C 30 sec]
- 72°C 3 min.
- 4°C, ∞
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Project name
