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定量引物
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: 产品保存液成分
主要成分：Na2HPO4、KH2PO4、NaCl和KCl、glucose
Q21: Can the morphology of this pseudovirus be captured by transmission electron microscopy? Can nucleic acid information be extracted?
The virus is granular and its morphology can normally be captured by a projection electron microscope. The nucleic acid sequences in viruses, except for the S protein sequence, are all nucleic acid sequences of pseudoviruses, not those of the novel coronavirus
Q22: Is the pseudovirus a soluble protein or particulate? If particulate, will adding it to LB or MSgg medium affect it?
Pseudoviruses are granular. Its influence in LB or MSgg medium was not tested
Q23: 如果假病毒颗粒中包裹有荧光蛋白是不是只有转染到细胞才能检测到荧光？
假病毒里只有荧光蛋白核酸序列，没有荧光蛋白，只有转染到细胞内才能发出荧光
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: 可否用投射电镜拍到该假病毒形貌，可否提取核酸信息
病毒是颗粒状的，正常是能用投射电镜拍到形貌；病毒里的核酸序列除了S蛋白序列之外，都是假病毒的核酸序列，不是新冠病毒的核酸序列
Q28: 假病毒是可溶的蛋白还是颗粒状的？如果是颗粒状的加入到LB或者MSgg培养基是否会对它有影响？
假病毒是颗粒状的；没有测试其在LB或MSgg培养基中的影响
Q29: 如果假病毒颗粒中包裹有荧光蛋白是不是只有转染到细胞才能检测到荧光？
假病毒里只有荧光蛋白核酸序列，没有荧光蛋白，只有转染到细胞才能检测到荧光，单独的假病毒不能检测到荧光
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: 荧光素酶的值
不同仪器、试剂盒不同，大概10的5-6次方
Q34: 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.
Q35: 我们可感染细胞的假病毒，用检测新冠的S蛋白试剂，可以检测出来的吧？
理论上是可以的，但是我们没检测过
Q36: 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.
Q37: 待感染细胞的每孔用量
1*10的4次方，培养24小时（约增长1.5倍左右，也就是2*10的4次方）
综合：待感染细胞和假病毒病毒的数量比推荐是：1:5~1:1
Q38: 我们的可感染细胞的假病毒，可以侵染Vero细胞么？
侵染Vero细胞没做过测试，目前我们做过测试过的只有经过ACE2过表达的293系列细胞，还有野生型的huh7细胞，但是huh7 的效率不高，还有一个bhk21细胞
Q39: 假毒是HIV骨架吗
是
Q40: 说明书中的mock 是什么
mock是空细胞
Q41: 假病毒里的荧光素酶用什么试剂盒激活？
我们用promega 的
Q42: 可感染细胞的假病毒是怎么构建的
是HIV骨架改造的，加了S蛋白包膜，载体质粒里有GFP和荧光素酶。常规慢病毒三个质粒=慢病毒的包装质粒+VSVG包膜外壳+载体质粒（含有目标基因）；可感染细胞的假病毒三个质粒=慢病毒的包装质粒+新冠S蛋白包膜外壳+载体质粒（含有GFP和荧光素酶）
Q43: 产品颜色
个别产品是偏粉红色，不影响使用
Q44: 荧光是单荧光还是双荧光
目前都是单荧光，萤火虫的
Q45: PCR quantitative primers
ZsGreen-QPCR-F：ACGAGTCCAAGTTCTACGGC；ZsGreen-QPCR-R：CGGGGATGATCTTCTCGCAG
Q46: 可感染细胞的假病毒目前情况
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）
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: 说明书中的mock 是什么
mock是空细胞
Q50: 有感染活性的假病毒内的S基因的3.8k基因序列或引物序列是什么
只提供相关参考文献，序列不对外公布
Q51: 可感染细胞的假病毒目前情况
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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