实验仪器使用技术

Zymolyase Dissolve in 10% glucose. Store aliquots (25 µl) at 20oC in common freezer for stock solutions Use once and dispose Comments: alternative is to dissolve in SCE, 40% gycerol, 20 mg/ml final. Can use multiple times. 上一篇：POSI BLOT SOLUTIONS 下一篇：10X TBE BUFFER

10X TBE BUFFER3 Liters324 g Tris base165g Boric acid120mls 0.5M EDTA (pH 8.0)autoclave for 20 min2 Liters216g Tris base110g Boric acid80mls 0.5M EDTA (pH 8.0)autoclave for 20 min1 Liters108g Tris base55g Boric acid40mls 0.5M EDTA (pH 8.0)autoclave for 20 min 上一篇：Zymolyase (3mg/ml) 下一篇：20X SS ...

20X SSPE BUFFER3 Liters525.9g NaCl sodium chloride82.8g NaH2PO4 sodium phosphate monobasic28.2g EDTA powder FW=372bring up to 2400mls with H2Oadd NaOH to pH 7.4 (~27mls/liter of 10N NaOH)autoclave for 20 min 2 Liters350.6g NaCl sodium chloride55.2g NaH2PO4 sodium phosphate monobas ...

0.5M EDTAFWMLpH8.0xg =292.250.50.undefined*93.06undefinedAdd_to_about_300mls_H2O_while_spinningph_to_8.0bring_up_to_500mlsfilter_sterilizenote~I_FW~I_372.24~J_M~I0.5~J_L~I_0.5~J_**93.06undefined_________上一篇：20X SSPE BUFFER 下一篇：CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL FOR YEAST

ChIP Protocol-Mechanical Breakage & FA Lysis BufferDay -2:o Start a 5ml overnight culture from a single colony.Day -1:o Start a larger overnight culture using part of the 5ml culture (subculture).Day One - Part One:1. Grow culture until it is in log phase: OD600=0.7 - 1.22. For each sample you collect, put t ...

Flow cytometry of yeast cells stained with PIShort Coursefill flow buffer tank, empty waste turn on FACSCalibu turn on computer double click on "Bob's FL3h" file on desktop (an error message will come up - hit "OK") under the Aquire menu: designate the file folder location and file name in Parameter Desc ...

Yeast sporulation (liquid)1. Grow a saturated yeast culture in YEPD.2. Take 0.2 ml and put into 5 ml sterile water. Spin 3-5'.3. Pour off water; resuspend cells in 5 ml 0.3% KOAc + 20 µg/ml Adenine + any required amino acids because of homozygous mutations.4. Incubate with aeration for 3-5 days at 23ºC (usually 4). Ob ...

FROGGINGFrog is 8x6 prong. keep in EtOH, flame, then dip in sterile dH2O before use. Dip & swirl into yeast diluted in microtiter plates. Place on dry plate 1-2sec, remove, will leave 1-2 ul.Serial dilutions (in steril water on YPD) -8 samples x 6 dilutions. Aim for initial dilution at ~OD 0.1 in 250 ul. 5X dilutions (do ...

Frozen PermanentsYEAST1) grow culture to stationary in YEPD2) Add 0.8ml of culture to 0.8ml of 30% glycerol in labelled freezer vials3) vortex and place at -80oC4) for strains with plasmids, pick a colony from a selective plate, grow to stationary in YEPD and freeze in glycerol. streak out on selective p ...

Protocol for Sending StrainsYou will need:Sterile pieces of foil (about 2.5 inches square). (Sterilize and store in a glass petri dish)1/4" sterile disks (from Difco, #1599-35, concentration disks 1/4", sterile blanks)Remove a disk from the bottle using a sterile toothpick. place on a piece of st ...

Removal of Counts : NaOH MethodUnwrap the membranes and stack them in a pile (Do not dry out the membranes completely). Place them individually into 1L 0.1 N NaOH 0.2 % SDS wash solution for exactly 20 min. Pour off the wash solution and neutralize with 1 L 0.5 M Tris (pH 7.5) 0.2 % SDS for 20 min. Place the membranes on 3 MM paper. Let ...

YAC transfer by Kar1 mating:Reference: Hugerat, Y., Spencer, F., Zenvirth, D. and Simchen, G. (1994). A versatile method for efficient YAC transfer between any two strains. Genomics 22, 108-1172. Spencer, F., Hugerat, Y., Simchen, G., Hurko, O., Connelly, C. and Hieter, P. (1994). Yeast kar1 mutants provide an ef ...

The light microscope, so called because it employs visible light to detect small objects, is probably the most well-known and well-used research tool in biology. Yet, many students and teachers are unaware of the full range of features that are available in light microscopes. Since the cost of an ins ...

1.实验进行前，无菌室及无菌操作台(laminarflow)以紫外灯照射30-60分钟灭菌，以70%ethanol擦拭无菌操作抬面，并开启无菌操作台风扇运转10分钟后，才开始实验操作。每次操作只处理一株细胞株，且即使培养基相同亦不共享培养基，以避免失误混淆或细胞间污染。实验完毕后，将实验物品带出工作台，以70%ethanol擦拭无菌操作抬面。操作间隔应让无菌操作台运转10分钟以上后，再进行下一个细胞株之操作。2.无菌 ...

Good sterile technique is the first and most important step in insuring consistent results when employing recombinant DNA and protein expression techniques. Sterile technique refers to procedures by which cultures may be manipulated without infecting the worker or contaminati ...

Each time the microscope is to be used it should be set up correctly to give a good image. Most often users forget to adjust the iris diaphragm and focus the condenser to give its optimum performance, this could result in objects in the preparation being undetected. There are many microscope set up procedu ...

由于电镜产生的电子束穿透能力很弱，必须把标本切成厚度小于0.1um以下的薄片才适用，这种薄片称为超薄切片。常用的超薄切片厚度是50-70nm。 在透射电镜的样品制备方法中，超薄切片技术是最基本、最常用的制备技术。超薄切片的制作过程基本上和石蜡切片相似，需要经过取材、固定、脱水、浸透、包埋聚合、切片及染色等步骤。 一、 取材的基本要求 组织从生物活体取下以后，如果不立即进行适当处理，会由于细胞内部各种酶的作用，出现细胞自溶现象。此外，还 ...

1、 湿热灭菌在同样的温度下，温热的杀菌效果比干热好，其原因有：①蛋白质凝固所需的温度与其含水量有关，含水量愈大，发生凝固所需的温度愈低。湿热灭菌的菌体蛋白质吸收水分，因较大同一温度的干热空气中易于凝固。②温热灭菌过程中蒸气放出大量潜热，加速提高湿度。因而湿热灭菌比干热所要温度低，如在同一温度下，则湿热灭菌所需时间比干热短。③湿热的穿透力比干热大，使深部也能达到灭菌温度，故湿热比干热收效好。 　　湿热灭菌法包括有：　　（1）煮沸法：煮沸 ...

实验 植物体内可溶性蛋白质含量的测定植物体内的可溶性蛋白质大多数是参与各种代谢的酶类，测其含量是了解植物体总代谢的一个重要指标。在研究每一种酶的作用时，常以比活（酶活力单位／mg蛋白）表示酶活力大小及酶制剂纯度。因此，测定植物体内可溶性蛋白质是研究酶活的一个重要项目。常用测定方法有Lowry法和考马斯亮蓝G-250染料结合法。本实验将分别介绍这两种方法。一、考马斯亮蓝法【原理】考马斯亮蓝G-250测定蛋白质含量属于染 ...

一、原理 研磨叶片得到的匀浆，经过滤、离心可制备叶绿体。叶绿体的被膜比较脆弱，分离叶绿体应在等渗的缓冲溶液中， 0 ～ 4 ℃温度下进行。叶绿体活力会随着离体时间延长而不断下降，因此，分离工作尽可能在短时间内完成。 二、仪器设备及试剂 （一）仪器设备 冰箱， 800 型离心机，扭力天平，显微镜， pH 计，研钵，量筒，移液管，离心管，脱脂纱布等。分离器皿都须在 0 ℃下预冷。 （二）试剂 1. 分离介质：含 0.33 mol/L 山梨醇， 50 mmol/L Tris-HCl （或 Tricine ） pH7.6 ， 5 mmol/L MgCl 2 , 10 mmol ...