{"authors":[{"id":"1930430222","orcid":"","email":"mcauchy@oswego.edu","name":"Michael Cauchy","affiliation":"State University of New York at Oswego"},{"id":"316694339","orcid":"","email":"akhan3@oswego.edu","name":"Ali A. Khan","affiliation":"State University of New York at Oswego"},{"id":"818027221","orcid":"","email":"yulia.artemenko@oswego.edu","name":"Yulia Artemenko","affiliation":"State University of New York at Oswego"},{"id":"739487002","orcid":"","email":"david.dunn@oswego.edu","name":"David A. Dunn","affiliation":"State University of New York at Oswego"}],"bricks":[{"abstract":"Custom-made, inexpensive and open-source electroporation apparatus for Dictyostelium discoideum with all functional modules and peripherals.","id":"719486065","functions":[{"id":"389996966","description":"","implementations":[{"id":"85007359","quantity":1,"type":"brick"},{"id":"1404619235","quantity":1,"type":"brick"},{"id":"725326220","quantity":1,"type":"part"},{"id":"320154449","quantity":1,"type":"part"},{"id":"1159884655","quantity":1,"type":"part"},{"id":"1775334525","quantity":1,"type":"part"},{"id":"1241573566","quantity":1,"type":"part"}]}],"authors":[],"files":[{"url":".\/.\/.\/usdata\/Complete1.png"}],"long_description":"Electroporation to introduce DNA into cells is a common technique in genetic engineering. Dictyostelium discoideum is a well-established eukaryotic model organism that is frequently genetically modified (transformed). We present here an electroporator capable of high efficiency D. discoideum transformation. Our electroporator consists of a high voltage power supply, pulse generator circuit and cell sample cuvette holder. The power supply is programmed to deliver desired voltage, which when activated by a double-pole, double-throw switch (DPDT below) charges the capacitor. Further switching allows the charged capacitor to deliver a precise, consistent exponential decay wave of the proper duration to the cuvette containing a D. discoideum cell and DNA mixture. The high-voltage circuit is housed in an electrically insulated enclosure - we chose a plastic electrical junction box with appropriate holes for switch mounting and cables to pass. A cuvette holder results from the 3D print of our stereolithography (.stl) CAD file. An electrophoresis chamber cover with cables was used as an adapter for banana plug\/jack connections to the power supply. Integral in development and testing was an inexpensive 1,000 volt digital multimeter. To provide hands-free monitoring of discharge circuit voltage, we connected the multimeter to the cuvette holder with 24 inch male-male banana test leads.","instructions":[{"name":"assembly","steps":[{"files":[],"description":"Plug electroporator power supply leads into gel electrophoresis lid.","components":[]},{"files":[],"description":"Plug electrophoresis lid leads into power supply.","components":[]},{"files":[],"description":"Plug electroporator cuvette leads into cuvette holder.","components":[]},{"files":[],"description":"Plug 24 inch male-male banana test leads into digital multimeter and the other end into the female end of stackable banana jacks of Step 2.","components":[]},{"files":[],"description":"Perform functions check: power on the digital multimeter, set to measure voltage (up to 1KV), and charge & discharge the capacitor with the pulse generator switch at lower voltage (e.g. 75V). If the device is functioning correctly, when the discharge begins the multimeter will display approximately 75V and go to ~50V volts in about 30sec.","components":[]}]}],"name":"Dictyostelium discoideum Electroporator","license":"CERN Open Hardware License","notes":""},{"abstract":"","id":"1920396730","functions":[{"id":"1767004534","description":"","implementations":[{"id":"108291712","quantity":1,"type":"part"}]}],"authors":[],"files":[],"long_description":"We chose to purchase a commercially available plastic junction box to enclose our pulse generator circuit. A ½ inch or 13mm hole was drilled into the enclosure cover for the DPDT and ? inch holes were marked and drilled in the enclosure which provided a nice fit for the plastic NM connectors and pass-through cables.","instructions":[{"name":"","steps":[]},{"name":"assembly","steps":[{"files":[{"url":".\/.\/.\/usdata\/Enclosure1.png"},{"url":".\/.\/.\/usdata\/Enclosure2.jpg"}],"description":"Drill two ? inch (22mm) holes in any side of the enclosure with centers at 34mm from the exterior bottom and horizontally 46mm apart on center. It might be helpful to label one hole \u201CPower supply\u201D and the other \u201CCuvette holder\u201D.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/Enclosure3.jpg"}],"description":"Drill ½ inch or 13mm hole in enclosure lid for switch mounting. Place the hole where hand-switching will be most comfortable.","components":[]}]}],"name":"Enclosure","license":"","notes":""},{"abstract":"","id":"85007359","functions":[{"id":"585941447","description":"","implementations":[{"id":"1653770866","quantity":2,"type":"part"},{"id":"1427392657","quantity":5,"type":"part"},{"id":"1149587839","quantity":1,"type":"part"},{"id":"1161249312","quantity":1,"type":"part"},{"id":"718879471","quantity":9,"type":"part"},{"id":"1267295566","quantity":1,"type":"part"},{"id":"1703802816","quantity":1,"type":"part"},{"id":"530776764","quantity":1,"type":"part"},{"id":"1472874281","quantity":4,"type":"part"},{"id":"884445350","quantity":6,"type":"part"},{"id":"310817272","quantity":6,"type":"part"},{"id":"853512084","quantity":4,"type":"part"},{"id":"89082890","quantity":1,"type":"part"},{"id":"1207683500","quantity":2,"type":"part"},{"id":"1920396730","quantity":1,"type":"brick"}]}],"authors":[],"files":[{"url":".\/.\/.\/usdata\/PGpreface2.png"},{"url":".\/.\/.\/usdata\/PGpreface1.jpg"}],"long_description":"Components and configurations of the electrical circuit are described here. For all connections, refer to the circuit diagram and photos of the assembled circuit.","instructions":[{"name":"assembly","steps":[{"files":[{"url":".\/.\/.\/usdata\/pg1.png"}],"description":"Cut 4 pieces of 18 AWG wire (for the switch) to 30cm lengths and strip approx. 5mm of bare wire from each end. Crimp one end of each wire to a ring terminal.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG2a.jpg"},{"url":".\/.\/.\/usdata\/PG2b.png"}],"description":"Attach ring terminals (with wire from step 1) to terminals 2, 3, 4, and 5 of the DPDT switch. Install switch into hole drilled into lid of case.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG3.jpg"}],"description":"Using the 4 pieces of 18 AWG wire with ring terminals from step 1, connect switch terminals 2 and 3 to the outboard ports of one terminal block and 4 and 5 to the outboard ports of another terminal block by inserting paired wires into outside wire cages of terminal blocks, then tighten terminal block screws.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG4.jpg"}],"description":"Clip resistor pins and bend at 90° angle to fit into breadboard terminals.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG5.jpg"}],"description":"Connect six 100K? resistors (R3-R8) in parallel into breadboards using jumper wires as needed to form connections.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG6.png"}],"description":"Connect two 1M? resistors (R1 and R2) in series into breadboards using jumper wires as needed to form connections.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG7.png"}],"description":"Connect four 1M? resistors (R9-R12) in series. A short jumper wire is also placed on the board as shown (lower right section of breadboard) that will become a link between terminal blocks from steps 2, 3, 9 and 10.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG8a.png"},{"url":".\/.\/.\/usdata\/PG8b.jpg"},{"url":".\/.\/.\/usdata\/PG8c.jpg"}],"description":"Ensure the capacitor is not charged before handling by measuring voltage across the terminals using the multimeter. Connect three 20cm flexible jumper wires to each pole of capacitor. Note that two leads from each capacitor pole are present. Wire terminals can be crimped around capacitor leads or soldered for establishing a more permanent connection. Attach capacitor leads to breadboard per circuit diagram.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG9a.jpg"},{"url":".\/.\/.\/usdata\/PG9b.jpg"}],"description":"Connect terminal blocks from switch leads to breadboard (see step 3).","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG10a.jpg"},{"url":".\/.\/.\/usdata\/PG10b.png"},{"url":".\/.\/.\/usdata\/PG10c.png"}],"description":"Cut 1 red and 1 black 60 in. test lead in half. Strip cut ends to leave approximately 5mm of bare wire. It might be helpful to tape the two pairs of one red and one black wire together every few centimeters. Pass stripped wire ends through NM plastic connectors about 10cm. Connect terminal blocks to the stripped ends by inserting paired wires into outside wire cages of terminal blocks, then tighten terminal block screws. Push NM plastic connectors into the cable pass-through holes of the enclosure.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG11.png"}],"description":"Pull the terminal block ends of the power supply and cuvette holder cables further through the pass-through holes with only sufficient play to insert the terminal blocks into the appropriate places on the breadboards.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG12.png"}],"description":"Adhere the breadboards and capacitor to the inside base of the enclosure. While we used 2-sided tape, glue could be used for a more permanent solution. Connect power supply and cuvette holder terminal blocks to breadboard.","components":[]},{"files":[{"url":".\/.\/.\/usdata\/PG13.png"}],"description":"Attach enclosure cover.","components":[]}]}],"name":"Pulse Generator","license":"","notes":""},{"abstract":"","id":"1404619235","functions":[{"id":"1060727575","description":"","implementations":[{"id":"2038737840","quantity":1,"type":"part"}]}],"authors":[],"files":[{"url":".\/.\/.\/usdata\/0-Cuvette_Holder.stl"},{"url":".\/.\/.\/usdata\/Final Cuvette Chamber.dwg"}],"long_description":"The cuvette holder provides the delivery vehicle for the electric field that causes electroporation. It is also notable that the cuvette holder was designed to keep inward pressure on the plug ends that contact the cuvette. It was an important consideration in our design to provide hands-free electroporation and the built-in spring-like nature of the resulting connection ensures constant contact - even when like charges build up on each plug and its corresponding cuvette contact - a force that opposes contact. It is also notable that we chose stackable banana plugs so that we could tandem a 1,000V multimeter to the cuvette holder - especially useful during pre-electroporation functions check.","instructions":[{"name":"","steps":[]},{"name":"assembly","steps":[{"files":[{"url":".\/.\/.\/usdata\/CuvHold1.png"},{"url":".\/.\/.\/usdata\/CuvHold2.jpg"},{"url":".\/.\/.\/usdata\/CuvHold3.png"}],"description":"3D print a Cuvette holder in PLA without support.","components":[]}]}],"name":"Cuvette Holder","license":"","notes":""}],"parts":[{"id":"108291712","files":[],"description":"","name":"Junction box","manufacturing_instruction":{"steps":[]},"supplier_part_num":"E987NR","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/www.lowes.com\/pd\/CARLON-Gray-Weatherproof-Pvc-Junction-Box\/1000975650","supplier":"Lowes"},{"id":"320154449","files":[],"description":"","name":"1KV DC Power supply (e.g., Bio-Rad PowerPac 1000)","manufacturing_instruction":{"steps":[]},"supplier_part_num":"","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"","supplier":"Bio-Rad (or other)"},{"id":"1653770866","files":[],"description":"","name":"400 Point solderless breadboard","manufacturing_instruction":{"steps":[]},"supplier_part_num":"2157693","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/www.jameco.com\/z\/BB-801-Goldtool-400-Point-Solderless-Breadboard-3-25-Lx2-125-W_2157693.html","supplier":"Jameco"},{"id":"1427392657","files":[],"description":"","name":"Male-Male fixed length jumper wires","manufacturing_instruction":{"steps":[]},"supplier_part_num":"2127718","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/www.jameco.com\/z\/WJW-70B-5-70-Piece-Jumper-Wire-Kit-22AWG-14-Lengths-10-Colors_2127718.html","supplier":"Jameco"},{"id":"725326220","files":[],"description":"","name":"Multimeter","manufacturing_instruction":{"steps":[]},"supplier_part_num":"61593","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/www.harborfreight.com\/11-function-digital-multimeter-with-audible-continuity-61593.html","supplier":"CEN-TECH"},{"id":"1149587839","files":[],"description":"","name":"Test Lead Wire 25 ft. red","manufacturing_instruction":{"steps":[]},"supplier_part_num":"Wire-TL-18-00-25","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/powerwerx.com\/test-lead-rubber-flexible-wire","supplier":"Powerwerx"},{"id":"1161249312","files":[],"description":"","name":"Test Lead Wire 25 ft. black","manufacturing_instruction":{"steps":[]},"supplier_part_num":"Wire-TL-18-00-25","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/powerwerx.com\/test-lead-rubber-flexible-wire","supplier":"Powerwerx"},{"id":"718879471","files":[],"description":"","name":"Male-Male flexible jumper wires","manufacturing_instruction":{"steps":[]},"supplier_part_num":"KIT-ZWx4","material_amount":"","manufacturer_part_num":"","material_unit":"NONE","url":"https:\/\/www.mouser.com\/ProductDetail\/BusBoard-Prototype-Systems\/KIT-ZWx4?qs=sGAEpiMZZMvh1pRuiUVjFZ0aAJXGPyuNDJiA5Z8ZR2Q%2FfInvTU3aKA%3D%3D","supplier":"Mouser"},{"id":"1267295566","files":[],"description":"","name":"Capacitor - 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