For CNP, a single layer of MEFs, MSCs, DCs, or HEK- 293T cells (~200,000 cells) was spread on a 1 cm x 1 cm 3D CNP silicon chip surface for overnight cell incubation. Plasmids pre-loaded in PBS buffer were injected into individual cells via nanochannels (~500 nm diameter and 5 μm spacing) using a 200 V electric field for 5 pulses at 10 ms/pulse with a 0.1 second interval. Various electroporation conditions were tested for best choice. BEP (Neon Transfection System, Thermo Fisher Scientific), Gene Gun (PDS-1000/He™ System, Bio-Rad) and Lipofectamine 2000 transfections were conducted according to manufacturers’ instructions. Ascl1/Brn2/Myt1l plasmids at a weight ratio of 2/1/1 and a concentration of 100 ng/mL in PBS buffer, according to the protocol in literature,40 were pre-mixed for transfection. Cell transfection of PTEN, miR-128, CD47, CDX-CD47, CREKA-CD47 and CD63-GFP plasmids followed the same procedure.
Gene gun pds 1000 he system
Manufactured by Bio-Rad
The Gene Gun (PDS-1000/He™ System) is a laboratory instrument used for the delivery of DNA, RNA, or other macromolecules into cells or tissues. It utilizes helium pressure to accelerate micro-carrier particles coated with the desired genetic material, allowing for the efficient introduction of genetic material into target cells.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using gene gun pds 1000 he system
1
Optimized Cellular Transfection via Nanochannels
2
Optimized Cellular Transfection via Nanochannels
3
Transient Transformation of Onion Epidermal Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The coding sequences of CML12, MLO4 and MLO4C were amplified by PCR using the primers XbaI-CML12/CML12-BamHI, XbaI-MLO4/MLO4-BamHI, and XbaI-MLO4C/MLO4-BamHI (Table S2 ). Applying the biolistic PDS-1000/He gene gun system (Bio-Rad, http://www.biorad.com/ , accessed on 11 July 2020), onion epidermal cells were transiently transformed. After cultivated on 1.5 MS culture medium (1.5% Agar, 1/2 MS) for 16–24 hours, the fluorescent signals were detected under a Leica confocal laser scanning microscope (LSM710, Carl Zeiss, http://www.zeiss.com/ , accessed on 21 March 2020) as described by Gookin et al [56 (link)].
4
Biolistic Transient Transformation of Onion Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The coding sequences of the genes were amplified by PCR using the primers listed in Supplementary Table S7 , and then cloned into the vectors pSPYNE-35S and pSPYCE-35S57 (link). AtPRD1 was also cloned in pSUPER1300. The resulting constructs were introduced into onion epidermal cells using a biolistic PDS-1000/He gene gun system (Bio-Rad, http://www.bio-rad.com ). Fluorescent signals were examined using a confocal laser scanning microscope (LSM710, Carl Zeiss, http://www.zeiss.com ) 16–24 h after transformation.
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
Revolutionizing how scientists
search and build protocols!