Archive 2019 - Wetlab Training

FAQ

Class I. Basics

1. Safety

2. Regulation

3. Protocols (Private)

4. How to design sample cohort

5. How to collect and manage samples

6. How to purify RNA from blood

7. How to check the quantity and quality of RNA

8. RNA storage

9. How to remove DNA contanimation

10. What is Spike-in

Class II. NGS I

1. How to do RNA-seq

2. How to check the quantity and quality of RNA-seq library

3. What is Smart-seq2 and Multiplex

Class III. NGS II

• 1. How to do Pico

• 2. How to improve Pico

  • 2.1 Replace pico kit by other independent reagents

  • 2.2 Introduce UMI & barcode into pico system

  • 2.3 Remove ribosomal cDNA by DASH/CRISPR method

Class IV. Validation

• 1. How to find and select candidates

  • 1.1 Differential Expression

  • 1.2 Alternative Splicing

• 2. How to find controls and references

• 3. How to do qRT-PCR

  • 3.1 How to select qPCR strategy

  • 3.2 How to reverse transcrip RNA to cDNA

  • 3.3 How to design qPCR plate

  • 3.4 How to minish qPCR error between replicats

  • 3.5 How to analysis qPCR results

• 4. How to do ddPCR

• 5. More validation methods

  • Alternative splicing: Agilent 2100 or gel electrophoresis

Class V. Patent and License

1. How to design different cohorts

2. How to prepare paperwork

3. How to prepare testing kit

APPENDIX

1. Common laboratory instruments

2. Frequently-used reagents and consumables

Framework