Log In Sign up
The largest database of trusted experimental protocols

Plko 1 puromycin lentiral vector

Manufactured by Merck Group
Visit Supplier

PLKO.1-puromycin lentiral vector is a laboratory tool used for genetic manipulation. It is a lentiviral vector that confers puromycin resistance, allowing for selection of transduced cells. The core function of this product is to enable the introduction and stable expression of genetic elements in target cell lines.

Automatically generated - may contain errors

Lab products found in correlation

Fl 335, by Santa Cruz Biotechnology (2 mentions) Ep769y, by Abcam (2 mentions)

Close spelling variants of this product

PLKO.1 vector PLKO vector PLKO.1 Puromycin

2 protocols using plko 1 puromycin lentiral vector

1

Validating MYB Knockdown Using shRNA Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
shRNA constructs cloned into the pLKO.1-puromycin lentiral vector were acquired from Sigma Mission shRNA library. Three shRNA targeted against MYB were obtained (Supplementary Table 9): MYB shRNA 1 (TRCN0000295917), MYB shRNA 2 (TRCN0000040058), and MYB shRNA 3 (TRCN0000040060). A scrambled sequence shRNA was used as a non-targeting control. Lentivirus for each shRNA was produced as described above. MYB knockdown was confirmed by western blot for three shRNA constructs in HEK293T cells as they do not require MYB for cellular fitness. HEK293T cells were transduced with shRNA lentivirus. Successful transductants were selected for 24 hours after lentivirus administration using 1 µg/mL puromycin. Western blots were performed using a MYB antibody (1:1000 dilution; EP769Y; Abcam) and GAPDH (1:2000 dilution; FL-335; Santa Cruz).
Canver M.C., Lessard S., Pinello L., Wu Y., Ilboudo Y., Stern E.N., Needleman A.J., Galactéros F., Brugnara C., Kutlar A., McKenzie C., Reid M., Chen D.D., Das P.P., Cole M., Zeng J., Kurita R., Nakamura Y., Yuan G.C., Lettre G., Bauer D.E, & Orkin S.H. (2017). Variant-aware saturating mutagenesis using multiple Cas9 nucleases identifies regulatory elements at trait-associated loci. Nature genetics, 49(4), 625-634.
+ Open protocol
+ Expand
2

Validating MYB Knockdown Using shRNA Constructs

Check if the same lab product or an alternative is used in the 5 most similar protocols
shRNA constructs cloned into the pLKO.1-puromycin lentiral vector were acquired from Sigma Mission shRNA library. Three shRNA targeted against MYB were obtained (Supplementary Table 9): MYB shRNA 1 (TRCN0000295917), MYB shRNA 2 (TRCN0000040058), and MYB shRNA 3 (TRCN0000040060). A scrambled sequence shRNA was used as a non-targeting control. Lentivirus for each shRNA was produced as described above. MYB knockdown was confirmed by western blot for three shRNA constructs in HEK293T cells as they do not require MYB for cellular fitness. HEK293T cells were transduced with shRNA lentivirus. Successful transductants were selected for 24 hours after lentivirus administration using 1 µg/mL puromycin. Western blots were performed using a MYB antibody (1:1000 dilution; EP769Y; Abcam) and GAPDH (1:2000 dilution; FL-335; Santa Cruz).
Canver M.C., Lessard S., Pinello L., Wu Y., Ilboudo Y., Stern E.N., Needleman A.J., Galactéros F., Brugnara C., Kutlar A., McKenzie C., Reid M., Chen D.D., Das P.P., Cole M., Zeng J., Kurita R., Nakamura Y., Yuan G.C., Lettre G., Bauer D.E, & Orkin S.H. (2017). Variant-aware saturating mutagenesis using multiple Cas9 nucleases identifies regulatory elements at trait-associated loci. Nature genetics, 49(4), 625-634.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!