Laboratory Protocols
Official step-by-step protocols for Solarbio products. Each protocol has been validated in Solarbio quality control laboratories using representative sample types and instrument configurations. Protocol times are estimates for experienced researchers; actual times may vary by sample type, batch size, and laboratory conditions.
Protocol Overview
This section contains 30+ validated laboratory protocols organized by application category. Each protocol includes:
- Complete materials checklist with recommended Solarbio product SKUs
- Step-by-step instructions with technical notes, incubation times, and centrifugation parameters
- Expected results and quality check criteria at intermediate stages
- Troubleshooting guidance for common failure modes
- Optimization notes for specific sample types and scaling
General Laboratory Guidelines
The following best practices apply to all protocols referenced in this documentation:
| Practice |
Recommendation |
Rationale |
| DNase/RNase-free consumables |
Use certified nuclease-free pipette tips and tubes for all nucleic acid work |
Prevents template degradation and false negatives |
| Cold chain maintenance |
Keep enzymes, master mixes, and competent cells on ice during handling |
Preserves enzymatic activity and transformation efficiency |
| Calibrated pipettes |
Pipettes should be calibrated every 6–12 months |
Ensures accurate reaction volumes, critical for qPCR reproducibility |
| Fresh aliquots |
Aliquot reagents subject to freeze-thaw damage (antibodies, enzymes, standards) |
Prevents activity loss from repeated freeze-thaw cycling |
| Water quality |
Use nuclease-free water (SKU R1600) for molecular biology; ultrapure water (18.2 MΩ·cm) for biochemical assays |
Avoids contaminating nucleases and metal ions that inhibit reactions |
| 70% ethanol sterilization |
Wipe all work surfaces and pipettes before RNA work |
Inactivates environmental RNases |
| Positive controls |
Include a positive control in every PCR, ELISA, and enzyme activity run |
Distinguishes reagent failure from sample-specific issues |
| Documentation |
Record lot numbers of all kit components for each experiment |
Essential for COA traceability and troubleshooting |
Molecular Biology Protocols
| Protocol |
Product Category |
Est. Lab Time |
Experience Level |
Sample Types |
| PCR Setup Guide |
PCR master mixes (PC1150–PC1165) |
1–2 h |
Beginner |
DNA, cDNA |
| qPCR Setup Guide |
qPCR master mixes (SR1110–SR1140) |
1.5–2 h |
Intermediate |
cDNA, gDNA |
| DNA Extraction Protocol |
Genomic DNA extraction (D1700) |
30–50 min |
Beginner |
Animal tissue, cells, blood |
| Gel Extraction Protocol |
Agarose gel DNA recovery (D1200) |
15–20 min |
Beginner |
Agarose gel slices |
| RNA Extraction Protocol |
Total RNA extraction (R1100, R1200) |
25–40 min |
Intermediate |
Tissue, cells, blood |
| Competent Cell Transformation |
Competent cells (C1100, C1180, C1300) |
2 h + overnight culture |
Intermediate |
Plasmid DNA, ligation products |
Molecular Biology Protocol Quick Reference
| Application |
DNA Template Required |
Typical Yield |
Downstream Use |
| Routine PCR |
10–100 ng gDNA or 1–50 ng cDNA |
5–20 μg PCR product |
Gel extraction, cloning |
| qPCR gene expression |
10–100 ng cDNA (per 20 μL reaction) |
Ct 20–32 (typical) |
Relative/absolute quantification |
| Genomic DNA extraction (tissue) |
20–50 mg tissue |
10–50 μg |
PCR, sequencing, genotyping |
| Gel extraction |
50–500 ng target band |
30–400 ng recovered DNA |
Ligation, sequencing |
| RNA extraction (tissue) |
30–50 mg tissue |
30–100 μg total RNA |
RT-PCR, RT-qPCR, RNA-seq |
| Competent cell transformation |
0.1–10 ng plasmid or 1–5 μL ligation |
10–5000 colonies |
Plasmid propagation, expression |
Protein and Cell Biology Protocols
| Protocol |
Product Category |
Est. Lab Time |
Key Applications |
| Western Blot Protocol |
Antibodies, ECL substrate, lysis buffers |
6–8 h (or 2 days with overnight antibody incubation) |
Protein expression analysis |
| ELISA Protocol |
ELISA kits (all targets, sandwich and competitive) |
3–5 h |
Cytokine quantification, biomarker detection |
| Cell Culture Protocol |
Cell culture reagents (D-PBS, trypsin, media, FBS) |
30 min per routine passage |
Adherent and suspension cell maintenance |
Key Recommendations for Protein Work
- Protein extraction: Always add protease inhibitor cocktail (P0100, 1:100) to RIPA lysis buffer (R0010) immediately before use. Work on ice throughout extraction to minimize proteolysis.
- Western blot transfer: For high-MW targets (>120 kDa), extend transfer time to 90 min at 300 mA or use 0.45 μm PVDF membrane (P1500). For low-MW targets (<20 kDa), reduce transfer to 30 min and use 0.2 μm PVDF.
- ELISA plate coating: If using in-house coating protocols (not Solarbio pre-coated plates), optimize coating concentration by checkerboard titration. Incubate coating buffer overnight at 4°C for maximum binding density.
Biochemical Assay Protocols
General Considerations for Enzyme Activity Assays
- Sample preparation: All tissue homogenates and cell lysates should be kept at 2–8°C or on ice during preparation. Perform centrifugation at 4°C to remove debris.
- Standard curve: Prepare a fresh standard curve for each assay run. Use at least 5 standard points plus a blank, and verify R² ≥ 0.99 before calculating sample concentrations.
- Reaction timing: For kinetic (rate) assays, record the initial absorbance (t = 0) immediately after adding the final reagent, then measure at the specified intervals. The reaction rate must be linear during the measurement window.
- Blank correction: Include a sample blank (sample + all reagents except substrate) to correct for endogenous absorbance. If a kit does not specify a sample blank, use deionized water as the reagent blank.
Protocol Selection by Application
| Research Application |
Recommended Protocol(s) |
Related Product Section |
| Gene expression analysis |
PCR Setup, qPCR Setup, RNA Extraction |
Molecular Biology |
| Protein expression analysis |
Western Blot, Protein Extraction |
Antibodies |
| Cytokine quantification |
ELISA Protocol |
ELISA Kits |
| Oxidative stress measurement |
Enzyme Activity Assay |
Oxidative Stress Assays |
| Cell viability screening |
Cell Culture Protocol |
Cell Proliferation |
| Gene cloning |
PCR Setup, Gel Extraction, Transformation |
Cloning Reagents |
Quality Control and Validation
Each protocol published in this section has been validated in Solarbio's QC laboratories under the following conditions:
- Replicates: Each protocol was tested with a minimum of 3 independent biological replicates
- Instruments: Bio-Rad T100 Thermal Cycler (PCR), Bio-Rad CFX96 (qPCR), Thermo Scientific NanoDrop One (nucleic acid quantification), BioTek Synergy H1 (absorbance/fluorescence), Bio-Rad ChemiDoc MP (gel imaging and western blot)
- Sample types: Protocols include representative data from at least 2 different sample matrices
- Pass criteria: PCR efficiency 90–105% (qPCR), R² ≥ 0.99 (standard curves), CV < 15% (technical replicates), recovery 90–110% (extraction protocols)
Protocol List
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