核心提示：10X TBE, per liter Sequencing dye -------------------10X TBE, per liter Sequencing dye ------------------- ---------------- 108g Tris base 80% formamide 55g boric acid 10mM NaOH 40ml 0.5M EDTA 1mM EDTA 0.1% xylene cyanol 10% APS 0.1% bromophenol blue ------------------- dissolve 0.1g ammonium persulfate in 10ml dH2O and store frozenGEL COMPOSITIONS per liter 6% 8% 20% ------------------------------------------------ acrylamide 57g 76g 190g bis acrylamide 3g 4g 10g urea 500g 500g 500g 10X TBE 100ml 100ml 100ml ddH2O 300ml 300ml 300ml ------------------------------------------------ Dissolve with mild heat, and adjust to 1 liter Filter, & store at room temperature in a closed container.1. Clean 1 side each of 2 glass plates , first with ddH2O, then EtOH, then 3 times with acetone. Scrub hard & work a small area at a time on the second acetone wash. The cleaner the plates, the easier it is to pour a gel without bubbles.2. Grease the corners of the bottom spacer & place it on the front plate. Grease the one end each of 2 side spacers & place them along the sides of the front plate, with the greased ends at the bottom. Fit the side spacers with the bottom spacer, & put extra grease at both spacer fittings. Lay the back plate onto the top plate, clean size down. Even-up the edges & clamp the edges together on the bottom & both sides with large clamps. Be sure the spacers are straight and fit together.3. To 100ml of the appropriate gel mixture, add 50ul TEMED and 400ul 10% APS, then stir. Turn the plates over & prop up the front with an inverted 100ml plastic beaker. Tip one edge down and place your thumb down on that edge at the top, thus sealing off the plate overhang. Pour the acrylamide solution slowly into this corner next to your thumb, allowing the solution to flow down the edge into the plates. When the solution reaches the bottom, straighten the plates and allow the solution to fill the plates from the bottom up. Don't allow the solution above the smaller plate to all drain in - add more before this happens. If you don't, bubbles will be pulled into the gel. If a bubble is formed , tap the glass to dislodge it. If it won't come up, tilt the plates vertically, and bang the plates until the bubble is removed, then carefully lower the plates & continue pouring.4. When the gel plates are full, pour more solution over the gel, then place the comb into the top of the plates through the acrylamide solution . Clamp the comb into place & allow the gel to solidify - about 1 hour.5. Remove the comb and the bottom spacer. Remove all but 2 clamps on each side, and clamp the gel into the gel box, being sure the corners at the top are sealed with plastic squares & vacuum grease . Using a syringe & TBE buffer, rinse the air out of the wells and the bottom of the gel.6. Heat the samples at 90C for 5 min, then cool on ice. While the samples are cooling, again rinse out the wells with buffer, being sure to rinse out all the urea-containing buffer from each well.7. Using a drawn-out glass capillary & mouth pipetter, load each sample onto the gel. Run the gel at up to 80 watts, with the fan on.8. When the gel is finished , take the plates out of the gel box, and CAREFULLY separate the plates with a metal spatula by prying from one bottom corner. 6% gels can be lifted from the glass with a piece of 3MM paper rubbed over the gel - then covered with saran wrap & dried on a gel dryer. 8-20% gels are lifted using a used piece of X-ray film rubbed over the gel, then covered with saran wrap.9. Autoradiography of hot samples is at room temperature; less hot samples are exposed with screens at -70C.
100% Pit Ventilation=Downflow of ventilating air完全的粪坑通风=换气风向下
Grouping of Fans in Ventilation Houses通风室里不同类别的风机
Single Fan Controller for multiple rooms可控多个房间的单独风扇控制器
Unique Duct system=uniform distribution of fresh air单独的通风管道系统=新鲜空气分布平均Integrated Heat Exchangers整合的热交换室
All Air Flows through the attic所有的空气流动都会经过阁楼
Multiple Room Capability—Common Pit多个房间——同一个粪坑
Age segregation by room--AIAO pigflow 每个房间对应着不同年龄的猪群——全进全出
Ventilation operating and Control is a 3 step process 换气操作有三个步骤：
Main Controller —constantly senses average room temperatures through the common pit and controls fan operation主控制器 ——通过粪坑感应器持续的感应猪舍平均温度和控制风扇运作
Manual hallway slides —set for minimum needs of each pig room manually. Normally adjust once a week. 手动走廊滑板——设定每个房间的最小通风量。通常每周调节一次
Automated Room Controller —senses individual room temperatures and adjusts each rooms ceiling inlets to provide more or less incoming fresh air to the room.自动房间控制器——感应每个房间的温度，然后通过调节屋顶的空气进口大小来提供适量的空气进入房间
Heat Exchangers recover heat exhausted from the pit fans that would otherwise be wasted. This dramatically decreases heating costs and improves air quality within the building.热交换室收集了废气当中原本会被浪费的热量。这大大降低能耗的同时，还改善了猪舍里空气的质量。
All the minimum air that enters the pig space is tempered in the hallway
弥散顶棚的好处ADVANTAGES OF PERFALU
< 铝制，更卫生，易于清洁，持久耐用Aluminium made for better hygiene, easy cleaning and durability.