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Secondary Antibodies
SAlexa Fluor-Conjugated
Other Fluorescent-Conjugated
Other-Conjugated
Colloidal Gold Reagent
Alkaline Phosphatase(AP)-Conjugated
Colloidal Gold-Conjugated
FITC-Conjugated
Biotin-Conjugated
Purified Secondary Antibodies
Allophycocyanin(APC)-Conjugated
CY Fluorescent-Conjugated
Phycoglobin(PE)-Conjugated
RBTIC-Conjugated
Peroxidase (HRP)-Conjugated
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TGF-β/Smad
Hippo
Antibody-drug Conjugate/ADC Related
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Inhibitors & Antagonists & Agonists
PI3K/Akt/mTOR
Endocrinology & Hormones
Protein Tyrosine Kinase/PTK
Angiogenesis
HIF
Immunology & Inflammation
Stem Cells
Apoptosis
Metabolic Enzyme & Protease
Autophagy
ER Stress
Cell Cycle
Cytoskeleton
DNA Damage & DNA Repai
Epigenetics
Ubiquitin
JAK/STAT
GPCR & G Protein
Neuronal Signal Pathway
MAPK
NF-κB
Membrane Transporter & Ion Channel
TGF-β/Smad
Hippo
Antibody-drug Conjugate/ADC Related
Others
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Pipette & Dispenser & Pipette Dispensing Instrument
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Other Small Instruments
Instruments
Pipette & Dispenser & Pipette Dispensing Instrument
Oscillating Mixing & Stirring & Dispersing Instrument
Temperature Control & Sampling & Grinding Crushing Instrument
Sterilization & Filtration Instruments
PCR Instrument & Light Glue Instrument
Spectrophotometer
Centrifuge
Shaker
Vacuum Pump
Other Small Instruments
FAQ—Solarbio Competent Cell
FAQ Documentation—Competent Cell
| Q01: | What should I do if no colonies grow after transformation with TOP10 in the experiment? |
| A01: | 1,Check if there are issues with the competent cells: perform a blank validation on a non-antibiotic plate. 2,Verify if the transformation was successful: try using a different plasmid. 3,Try a 42°C heat shock for 45s or 60s. |
| Q02: | There is no resistance effect at high concentration, which is not consistent with the expected result. I used to use hygromycin from Shanghai sangon company, the same density has no effect! |
| A02: | The potency of antibiotics varies among different manufacturers. We conducted a bacteriostatic test using E.coli and found that 150μg/ml can inhibit bacterial growth, while lower concentrations have no effect. For fungi, a higher concentration than for bacteria is required. (For plant cells, the effective concentration ranges from 20–200μg/ml; for bacteria, it is 20–200μg/ml; for fungi, it is 200μg–1 mg/ml). We recommend increasing the dosage. |
| Q03: | The DH5α competent cells with number C1100 did not grow spots after we connected the products for transformation and plating. Three people did different transformation and all got the same result. |
| A03: | 1,Please confirm if the ligation conditions are appropriate. Note that when shaking the ligation product, do not add medium containing antibiotics. 2,Try transforming with a plasmid to see if colonies still do not grow, or perform a blank control. |
| Q04: | What are the differences between the various competent cells offered by your company? |
| A04: | 1,C1100 DH5 Strain: A cloning strain used for molecular cloning and plasmid extraction. It can be used for blue-white screening. 2,C1200 TOP10 Strain: A cloning strain also used for molecular cloning and plasmid extraction. Suitable for blue-white screening. This strain is ideal for efficient DNA cloning and plasmid amplification, ensuring stable genetic transmission of high-copy plasmids. 3,C1300 JM109 Strain: A cloning strain for molecular cloning. It is a suppressor strain with a recombinant deficiency that supports the growth of vectors with amber mutations. It modifies transfected DNA without restriction. 4,C1400 BL21(DE3) Strain: An expression strain used for expressing non-toxic proteins. This strain is suitable for efficiently expressing genes cloned into expression vectors containing the T7 phage promoter. It is suitable for the expression of non-toxic proteins. 5,C1500 BL21(DE3)pLysS Strain: An expression strain capable of expressing both toxic and non-toxic proteins. The pLysS strain contains the pLysS plasmid, conferring chloramphenicol resistance. The pLysS plasmid carries a gene that expresses T7 lysozyme, which reduces background expression levels of the gene of interest without interfering with the expression of the target protein. It is suitable for the expression of both toxic and non-toxic proteins. |
| Q05: | In Blue and white screening, shoule I choose the blue colonies or white colonies? |
| A05: | On selection media containing X-gal and IPTG, transformants carrying vector DNA appear as blue colonies, while transformants with recombinant plasmids containing inserted fragments appear as white colonies. To fully develop the color reaction, the plates can be incubated at 37°C followed by placement in a refrigerator for 3-4 hours, making the blue colonies more pronounced. |
| Q06: | What are the recommended concentrations of commonly used antibiotics? |
| A06: | Ampicillin (Amp): 100μg/mL (for bacteria); Kanamycin (Kan): 10-50μg/mL (for bacteria); Streptomycin (Str): 10-100μg/mL; Rifampicin (Rif): 50-100μg/mL; Gentamicin: 10-50μg/mL; Amphotericin: 0.25μg/mL |
| Q07: | Why is the efficiency of connection and transformation low? |
| A07: | 1,The ligation system is not suitable; 2, Blunt-ended ligation is difficult and requires overnight incubation at 4°C with gentle handling; 3,Shorten the heat shock time to 45s; 4,Check if the enzyme is suitable (e.g., pfu amplification may require separate addition of a polyA tail). |
| Q08: | Why do other bacteria grow after transformation? |
| A08: | If satellite colonies emerge around individual colonies, it indicates that the incubation time was prolonged, resulting in strain aging. This has minimal impact on subsequent experiments and can be rectified by reactivation. If colonies of similar sizes are observed, it suggests contamination during the operation. |
| Q09: | Why do only a few colonies grow after transformation? |
| A09: | 1, If the transformed product is a ligation product or a mutant product, this is considered normal. If plasmids are used, attention should be paid to potential issues in the manipulation process. 2, Some plasmids inherently have low transformation efficiency; trying a different competent cell type may improve the outcome. |
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