Myc CRISPRa sgRNA lentivector (set of three targets)(Mouse)

0

Question

How should I store my lentivirus?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

The lentivirus should be aliquoted into smaller working volumes and then stored at -80°C. Lentivirus is sensitive to storage temperature and to freeze/thaws. It can lose up to 5% or more activity with each freeze/thaw. When stored properly, viral stocks of an appropriate titer should be suitable for use for up to one year. After long-term storage, we recommend re-titering your viral stocks before use.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What generation does abm use for their lentiviral transfer vectors?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

abm lentiviral transfer vectors use the third generation lentivirus system. It is based on the inactivated HIV genome. Note that our lentivirus packaging plasmids cannot be integrated into the host and are transiently expressed.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

How do I calculate the Multiplicity of Infection (MOI)?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

MOI (Multiplicity of Infection) refers to the number of viral particles per cell used in the infection, e.g. an MOI of 5 indicates that there are five infectious units (IU) or transducing units (TU) for every cell. MOI is determined by calculating the numbers of viral particles added per well then divide this number by the number of cells seeded into the well. We also recommend transducing the cells with a range of MOIs as different cell types may require different MOIs for successful transduction.

MOI = Product Titer (IU/ml) x Virus Volume (ml) / Total Cell Number

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What concentration of puromycin should I use for cell selection following lentivirus transduction?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

The concentration must be optimized for each cell type. Typical selection amounts are around 0.1 - 0.5µg per ml.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What is the optimal cell density for lentivirus transduction?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

Optimal cell density for lentiviral transduction is generally 50-70% confluent for most cell types. Certain cell lines may require the optimal cell density to be determined experimentally.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

Are abm’s vectors high or low copy? What is the expected yield from a plasmid extraction prep?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

These are medium-high copy plasmids and should be propagated in a cloning E. coli strain such as DH5α. Typical yields from a 250ml culture is 300-500µg plasmid DNA.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

How are abm’s vectors shipped and what buffer are they supplied in?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

Standard delivery of abm’s vectors are in liquid format supplied in TE buffer (10mM Tris, 1mM EDTA, pH 8.0). Our vectors can also be ordered and delivered in agar stabs for an additional fee.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What concentration are abm’s vectors supplied as?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

Typically vectors are supplied at 100ng/µl.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

How should I store abm’s vectors?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

We recommend storing abm’s vectors at -20°C and avoiding repeated freeze/thaws as this may affect plasmid integrity and cloning efficiency.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What is MOI and how do I know which MOI to use?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

MOI stands for multiplicity of infection. Theoretically, an MOI of 1 will provide 1 virus particle for each cell on a plate, while an MOI of 10 represents ten virus particles per cell. However, several factors can influence the optimal MOI including cell line, cell type, transduction efficiency and gene of interest. We recommend first establishing an optimal MOI for each cell line. This can be done using a range of MOIs (0, 0.5, 1, 2, 5, 10, 50) to determine the MOI required to obtain optimal gene expression

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What buffer is abm’s lentivirus provided in?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

10^7 IU/ml = DMEM
10^8, 10^9, 10^10 IU/ml = 1X PBS (pH 7.4) with 2% PEG6000

ABM Scientific Support

Answered on Jan 30 2026

0

Question

How do I transform abm’s plasmids? What bacterial strain should I use?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

A general Plasmid Amplification Protocol can be found here.

We recommend transforming abm’s plasmids into our ProClone™ Competent Cells (Cat. No. E003). The ProClone™ Competent Cells are high-efficiency chemically competent DH5α (E. coli) cells ideal for routine plasmid amplification due to its high yield, rapid growth, and high transformation efficiency, with recA⁻ and endA⁻ mutations that ensure plasmid stability and high DNA quality. Other common compatible cloning strains include TOP10 (DH10B derivatives).

ABM Scientific Support

Answered on Jan 30 2026

0

Question

How do I revive an agar stab?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

1. Use a sterile loop or pipette tip to scrape a small amount of cells from the agar stab. Only a tiny amount is needed as it contains live E. coli.
2. Gently streak the cells onto an agar plate containing the correct antibiotic selection in order to achieve single isolated colonies.
3. Place the plate “agar side up” and incubate at 37°C overnight.
4. Select a single colony from the plate for downstream applications, such as an overnight broth culture and grow E. coli to late exponential-early stationary phase. 4a. The broth culture can be subjected to miniprep plasmid extraction. We recommend using abm’s Column-Pure Plasmid Miniprep Kit (Cat. No. G4003). 4b. The broth culture can be used to prepare a glycerol stock for long-term storage. Mix culture with sterile glycerol to a final concentration of 15% and store at -80°C.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

I packaged abm’s lentivector into virus, performed transduction and antibiotic selection. Following selection all my cells died. What should I do?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

We recommend first checking the following parameters:
1. Cell density. If density is too low, this can also result in cell death.
We recommend the following experiments:
Initial Experiment
•Transduce lentivirus into the 293T cell line - one of the easiest cell lines to transduce. If the transduced cells survive selection, then troubleshooting should focus on post-virus production steps. If the transduced cells didn't survive selection, the troubleshooting should focus on virus packaging steps.
Virus Packaging Troubleshooting
•Attempt lentivirus packaging using a GFP expressing control vector. If packaging is successful with the control, then repeat packing of the lentivirus in question with a GFP batch control to ensure no errors with the first production. If no GFP signal is observed after transfection or ~72h after a test transduction of a GFP control lentivirus on 293T cells, then the packaging process needs to be assessed. Packaging process assessment includes reagent quality, cell health, and packaging protocol details.
Post-Virus Production Troubleshooting
•Assess if the correct antibiotic concentration is used. This can be done by performing a drug-killing curve. To set up a drug-killing curve, we recommend using the same culture size and seeding density for your actual selection, and adding a different puromycin concentration to each sample, with the range between 0-1 ug/ml. If the cells are not killed at the 1ug/ml, you may try increasing the range higher concentrations. It is important to identify the concentration that results in >95% cell death in 1-4 days to establish the minimal concentration to use for the selection process.
•Assess if the correct MOI is used to transduce the target cells. This can be done by using a GFP control lentivirus to transduce the target cells at a range around literature-recommended MOIs to determine the optimal MOI for transduction.
•Assess if a transduction enhancer such as the ViralEntry Transduction Enhancer (Cat.No. G515) is necessary. Some cell lines are more difficult to transduce than others and by using a transduction enhancer can lead to increased target cell permeability.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

I transduced my cell line using abm’s virus. Afterwards my cells exhibited a different morphology and/or died. Why?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

1. This often happens to primary cells which are very sensitive to culture medium conditions. Try to use a higher titer virus in order to limit the volume of exogenous media added to your cells.
2. Virus preparation might be contaminated with bacteria. Use a 0.25µm syringe filter to clear the virus preparation and repeat transduction following strict sterile technique.
3. Cell line might be contaminated with mycoplasm. This type of contamination is often not immediately obvious. The contamination effect is amplified after virus transduction. Repeat transduction with fresh cells.
4. The virus preparation may contain some cell debris which could be mistaken as a change in cell morphology. Normally this issue will disappear 3-5 days after virus transduction.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

I am interested in one of abm’s vectors. Where can I find the corresponding control?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

A complete list of controls can be found on abm’s Control Vectors and Viruses page.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What troubleshooting steps do you recommend if plasmid amplification fails?

ABM community

Verified customer

Asked on Jan 29 2026

0

Answer

Verify plasmid quality: Confirm the plasmid is intact, at sufficient concentration, and free of contaminants (e.g., salts, ethanol, nucleases).
Check the host strain: Use a suitable cloning strain (e.g., recA⁻/endA⁻ such as DH5α) and confirm cells are competent and not expired.
Confirm selection conditions: Make sure the correct antibiotic is used at the proper concentration and that plates/media are fresh.
Review transformation conditions: Ensure the transformation method (chemical or electrocompetent) and recovery time are appropriate.
Assess plasmid compatibility: Large, low-copy, toxic, or unstable plasmids may require specialized strains or growth conditions.
Inspect growth conditions: Verify incubation temperature, shaking speed, and culture time.
Check plasmid sequence/features: Look for toxic genes, strong promoters, or repeats that may reduce stability.

ABM Scientific Support

Answered on Jan 29 2026

0

Question

What are CRISPR activation & repression vectors and how do they work?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

These constructs are a collection of lentiviral vectors that express sgRNAs targeting the upstream 5′ UTR of a specific gene. When combined with dCas9-VPR, they mediate targeted gene activation by recruiting transcriptional activators. In contrast, pairing the sgRNAs with dCas9-KRAB results in targeted gene repression through KRAB-mediated transcriptional silencing.

ABM Scientific Support

Answered on Jan 30 2026

0

Question

What type of Cas9 is compatible with the CRISPR Activation & Repression vectors?

ABM community

Verified customer

Asked on Jan 30 2026

0

Answer

We recommend using dCas9-VPR for gene activation experiments and dCas9-KRAB for gene repression experiments.

ABM Scientific Support

Answered on Jan 30 2026

Myc CRISPRa sgRNA lentivector (set of three targets)(Mouse)

Retired Cat.No.

Product Name

Unit

Description

Gene Name/Gene ID

Gene Full Name

Alias

Accession Number

Species

System

Target Sequence

Promoter

Storage Condition

QC

Disclaimer

Caution