Q1: If pseudovirus particles encapsulate a fluorescent protein, can fluorescence only be detected after transfection into cells?
In pseudoviruses, there are only nucleic acid sequences of fluorescent proteins, but no fluorescent proteins. Fluorescence can only be detected when transfected into cells. Fluorescence cannot be detected in a single pseudovirus.
Q2: Virus particle size
About 100 to 120 nanometers.
Q3: Shelf life at 4°C
The impact is not significant within a short period of 12 hours, but if it lasts for a long time, the activity will decrease or even become unusable. Store at -80 degrees Celsius and avoid multiple freeze-thaw cycles.
Q4: Titer
Gradient dilution method, greater than 1 × 10 to the power of 7. We use the gradient dilution method. We use the same number of cells. For example, if the number of cells is all 1E+5, if 10ul of the virus is added and all the cells are fully infected, it is overinfection. If 1ul is added and 10% of the cells are infected, then 1ul of the virus has infected 1E+4 cells. Therefore, 1ml can infect 1E+7 cells. It's roughly calculated like this.
Q5: Luciferase value
Different instruments and reagent kits vary, approximately 10 to the power of 5 to 6.
Q6: In neutralization assays, how much reduction is considered effective?
This is hard to define. A 50-70% ratio can be considered to have a relatively good effect.
Q7: Can our infectable pseudovirus be detected using an S protein detection reagent for COVID-19?
Theoretically, it's possible, but we haven't tested it.
Q8: Dosage for infectable cells
The amount of pseudovirus that can infect cells in each well of a 96-well plate is 2-10 μ l, and the pseudovirus concentration is 1-10 to the power of 7, with 1 μ l being 10 to the power of 4.
Q9: Amount per well for cells to be infected
At 110 to the power of 4, cultivate for 24 hours (about 1.5 times the growth, that is, 210 to the power of 4). Overall: The recommended ratio of the number of cells to be infected and the pseudovirus is 1:5 to 1:1.
Q10: Can our infectable pseudovirus infect Vero cells?
Vero cells have not been tested for infection. Currently, the only cells we have tested are 293 series cells with ACE2 overexpression, wild-type huh7 cells, but the efficiency of huh7 is not high, and there is also a bhk21 cell.
Q11: Is the pseudovirus based on an HIV backbone?
Yes.
Q12: What does "mock" mean in the instructions?
mock is an empty cell.
Q13: What kit is used to activate the luciferase in the pseudovirus?
We use promega.
Q14: How is the infectable pseudovirus constructed?
It is modified from the HIV skeleton, with an S protein envelope added, and the carrier plasmid contains GFP and luciferase. The three plasmids of a regular lentivirus = the packaging plasmid of the lentivirus +VSVG envelope shell + vector plasmid (containing the target gene). The three plasmids of pseudoviruses that can infect cells = lentivirus packaging plasmid + SARS-CoV-2 S protein envelope shell + carrier plasmid (containing GFP and luciferase).
Q15: Product color
Some products are slightly pink in color, but this does not affect their use.
Q16: Is it single or dual fluorescence?
At present, they are all monofluorescent fireflies.
Q17: PCR quantitative primers
ZsGreen-QPCR-F：ACGAGTCCAAGTTCTACGGC ZsGreen-QPCR-R：CGGGGATGATCTTCTCGCAG。
Q18: What is the 3.8k gene sequence or primer sequence for the S gene in the infectious pseudovirus?
Only relevant references are provided. The sequence is not disclosed to the public.
Q19: What is the current status of the infectable pseudovirus?
At present, the pseudoviruses that can infect cells mainly consist of two types: the HIV skeleton and the VSV skeleton. Both types of viruses can be used in cell experiments and animal experiments. The VSV skeleton pseudovirus is slightly more efficient in animal experiments. Our pseudovirus is based on the HIV skeleton. The three plasmids of the HIV skeleton pseudovirus = the packaging plasmid of lentivirus (HIV) + the encapsulation shell of the S protein of COVID-19 + the vector plasmid (containing GFp and luciferase). The three plasmids of the VSV skeleton pseudovirus = the packaging plasmid of the vesicular stomatitis virus vector (VSV) + the envelope shell of the SARS-CoV-2 protein + the vector plasmid (containing GFp or luciferase). Animal experiments with pseudoviruses: First, adenovirus is used to induce overexpression of ace2 cells in some tissues or organs of animals, and then pseudoviruses that can infect cells are used for infection experiments.
Q20: Ingredients of the product preservation solution
Ingredients of the product preservation solution
Q21: Can the morphology of this pseudovirus be captured by transmission electron microscopy? Can nucleic acid information be extracted?
Can the morphology of this pseudovirus be captured by transmission electron microscopy? Can nucleic acid information be extracted?
Q22: Is the pseudovirus a soluble protein or particulate? If particulate, will adding it to LB or MSgg medium affect it?
Is the pseudovirus a soluble protein or particulate? If particulate, will adding it to LB or MSgg medium affect it?
Q23: If the pseudovirus particles are encapsulated with fluorescent proteins, can the fluorescence only be detected when transfected into cells?
If the pseudovirus particles are encapsulated with fluorescent proteins, can the fluorescence only be detected when transfected into cells?
Q24: Can a positive control be provided?
Positive controls can be provided. They have been prepared and cost 150 yuan per copy
Q25: Can it be used for IC50 determination?
It can be done
Q26: Composition of the product storage solution
Because it is the solvent of the virus preservation solution, there may be trace amounts of serum, DMEM medium residue, and some impurity proteins from cells remaining in the virus, all of which cannot be completely removed
Q27: Can the morphology of the pseudovirus be captured by a projection electron microscope? Can nucleic acid information be extracted
Can the morphology of the pseudovirus be captured by a projection electron microscope? Can nucleic acid information be extracted
Q28: Is the pseudovirus soluble protein or granular? If it is in granular form and added to LB or MSgg medium, will it have any impact on it?
Is the pseudovirus soluble protein or granular? If it is in granular form and added to LB or MSgg medium, will it have any impact on it?
Q29: If the pseudovirus particles are encapsulated with fluorescent proteins, can the fluorescence only be detected when transfected into cells?
If the pseudovirus particles are encapsulated with fluorescent proteins, can the fluorescence only be detected when transfected into cells?
Q30: What is the virus particle size?
About 100 to 120 nanometers.
Q31: What is the shelf life when stored at 4°C?
The impact is not significant within a short period of 12 hours, but if it lasts for a long time, the activity will decrease or even become unusable. Store at -80 degrees Celsius and avoid multiple freeze-thaw cycles.
Q32: What is the titer?
Gradient dilution method, greater than 1 × 10 to the power of 7. We use the gradient dilution method. We use the same number of cells. For example, if the number of cells is all 1E+5, if 10ul of the virus is added and all the cells are fully infected, it is overinfection. If 1ul is added and 10% of the cells are infected, then 1ul of the virus has infected 1E+4 cells. Therefore, 1ml can infect 1E+7 cells. It's roughly calculated like this.
Q33: The value of luciferase
The value of luciferase
Q34: In neutralization assays, how much reduction is considered effective?
In neutralization assays, how much reduction is considered effective?
Q35: The pseudovirus that can infect cells can be detected by using the S protein reagent for detecting COVID-19, right?
The pseudovirus that can infect cells can be detected by using the S protein reagent for detecting COVID-19, right?
Q36: Dosage for infectable cells
Dosage for infectable cells
Q37: The dosage per well of the cells to be infected
The dosage per well of the cells to be infected
Q38: Can our pseudovirus that can infect cells infect Vero cells?
Can our pseudovirus that can infect cells infect Vero cells?
Q39: Is the pseudovirus the skeleton of HIV
Is the pseudovirus the skeleton of HIV
Q40: What is "mock" in the instruction manual
What is "mock" in the instruction manual
Q41: What kit is used to activate the luciferase in pseudoviruses?
What kit is used to activate the luciferase in pseudoviruses?
Q42: How are pseudoviruses that can infect cells constructed
How are pseudoviruses that can infect cells constructed
Q43: Product color
Product color
Q44: Is the fluorescence single fluorescence or double fluorescence
Is the fluorescence single fluorescence or double fluorescence
Q45: PCR quantitative primers
PCR quantitative primers
Q46: The current situation of pseudoviruses that can infect cells
The current situation of pseudoviruses that can infect cells
Q47: Can the cell-infecting COVID-19 pseudovirus infect mice?
Our pseudovirus of the novel coronavirus has been tested on animals, but the results were not good and it could not be detected. It is not recommended for infecting mice.
Q48: Can it be used in a laboratory cell culture room? Any harm to the laboratory and personnel?
The pseudovirus contains no COVID-19 nucleic acid at all. It only has S protein on its surface and is not replicable. It can be used in laboratory cell rooms and is not pathogenic. What is written in our instruction manual is relatively conservative.
Q49: What is "mock" in the instruction manual
mock是空细胞
Q50: What is the 3.8k gene sequence or primer sequence of the S gene in a pseudovirus with infectious activity
只提供相关参考文献，序列不对外公布
Q51: The current situation of pseudoviruses that can infect cells
可感染细胞的假病毒目前主要由HIV骨架和VSV骨架两种，两种病毒都可以用于细胞实验和动物实验，VSV骨架假病毒做动物实验效率会略好一些。我们的假病毒是HIV骨架的。
HIV骨架假病毒三个质粒=慢病毒(HIV)的包装质粒+新冠S蛋白包膜外壳+载体质粒（含有GFp和荧光素酶）
VSV骨架假病毒三个质粒=水泡性口炎病毒载体（VSV）的包装质粒+新冠S蛋白包膜外壳+载体质粒（含有GFp或荧光素酶）
假病毒的动物实验：先用腺病毒让动物的部分组织或器官有ace2细胞的过表达，在用可感染细胞的假病毒进行侵染实验
| 类别 |
产品情况 |
货号 |
名称 |
| 原始株 |
有 |
BDAA0026 |
FNV-SARS-CoV-2-S（可感染细胞） |
| Alpha |
无 |
|
|
| Beta |
只有三个点突变株（E484K、K417N、N501） |
BDAA0084 |
FNV-SARS-CoV-2-S(可感染细胞)(E484K、K417N、N501Y) |
| Gamma |
只有三个点突变株（E484K、K417N、N501） |
BDAA0084 |
FNV-SARS-CoV-2-S(可感染细胞)(E484K、K417N、N501Y) |
| Delta |
有 |
BDAA0115 |
FNV-SARS-CoV-2-S(可感染细胞)(B.1.617.2)(delta) |
| Lambda |
无 |
|
|
| Omicron |
有 |
BDAA0117 |
FNV-SARS-CoV-2-S(可感染细胞)(B.1.1.529)(Omicron) |
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