其他模型

Grow 100 ml yeast cells in desired media overnight to an A600 of ~1.0 (0.6 to 1.2 works well). For growth in minimal media 1 ml of a saturated overnight culture in minimal media (Synthetic dextrose (S ...

Procedure pick one colony inoculate in 3 ml of the appropriate media grow at 30° overnight pellet the cells (5 min 5000g) wash 1X in sterile ddH2O re- suspend the pellet in 200 µl EthOH (optional: +2 ...

This method gives yeast nuclei which look nearly purified microscopically. Nuclei isolated in this way do not give active transcription extracts when the nuclei are salt extracted and the resulting pr ...

This is the preferred method for yeast RNA preparation use Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fresh YPD media and grow cells at 30 degrees overnigh ...

AbstractWhen more than one bait will be used to screen a single library significant time and resources can be saved by performing the interactor hunt by interaction mating. In this protocol one strain ...

Express fusion proteins as per the GST-fused protocol up to Step 7 (Day 3). All steps in protein purification should be done at 4° C unless otherwise stated.Pre-wash 10-20 ml of New England Biolabs am ...

Protocol: Fast yeast transformationAdd 50 µl carrier DNA to a 1.5 ml tube. scrap cells from plate and add to the carrier DNA. Add in the following order: 1-2 µl DNA-plasmid (up to 30-50 µl) 240 µl PEG ...

Day 0 Make sure you have the necessary solutions (instructions for how to make them can be found here): Single-stranded carrier DNA PEG 3350 50% w/vol 1.0 M LiAc Make sure you have enough plasmid DNA ...

Short term storageYeast cultures are stable for 1-2 weeks when refrigerated. Petri dishes should be sealed or in plastic bags. Medium term storageYeast can be preserved for up to a year on YPD agar me ...

Grow cells Harvest Spin down Wash with PBS Spin down enough cells to collect a 50-100 µL cell pellet in a FastPrep tube Prepare lysis buffer (beforehand) and chill on ice PBSMT: 2.5 mM MgCl2 3 mM KCl ...

Pad Preparation1. Microwave 2% agarose (mix of low-melt and normal to taste) in Thorn media (see below). (If you have used different percentages of agarose pads please let us know bruno) 2. Apply 2mL ...

OverviewBlotting for large V5-tagged proteins in S. cerevisiae MaterialsY-PER (Pierce) Halt EDTA-free Protease Inhibitor (Pierce) NuPAGE Novex Bis-Tris 4-12% mini gels (Invitrogen NP0321BOX) Prestaine ...

MaterialsYPD plates YPD 1 M sorbitol 182 g/l (Sigma S7547) 2 M sorbitol 36.4 g/100 ml SCE (per liter) 1 M sorbitol (182 g) 100 mM citric acid trisodium salt dihydrate (29.4 g) 10 mM EDTA 20 ml of 500 ...

Protocolgrow up yeast culture to appropriate density (near saturation) spin 1.5 mls of culture for 1 min in microfuge and aspirate off supernatant resuspend pellet in 200 ul breaking buffer wear glove ...

INTRODUCTION The Drosophila larval brain is a well-established model for investigating the role of stem cells in development. Neuroblasts (neural stem cells) must be competent to generate many thousan ...

SOS信号转导途径在植物离子平衡和耐盐中非常重要。SOS模型认为高Na+引起了胞内自由Ca2+的升高，激活了Ca2+结合蛋白编码的SOS3的表达，影响到下游的反应。SOS3激活了相连的SOS2（丝氨酸/苏氨酸蛋白激酶），SOS2/SOS3复合体调节盐忍耐因子编码的SOS1（质膜Na+/H+反向转运体）的表达水平，从而促使胞内过多的Na+排出体外。这其中涉及到K+的变化，Na+和K+平衡的破坏可能是 ...

Single-Cell Electroporation in XenopusXue Feng Liu and Kurt Haas INTRODUCTIONSingle-cell electroporation (SCE) is a versatile technique for delivering electrically charged macromolecules including DNA ...

小鼠(T)酶联免疫分析（ELISA）试剂盒使用说明书本试剂仅供研究使用目的：本试剂盒用于测定小鼠血清，血浆及相关液体样本中*~AT~B含量。实验原理：本试剂盒应用双抗体夹心法测定标本中小鼠*~AT~B水平。用纯化的小鼠*~AT~B抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入*~AT~B，再与HRP标记的*~AT~B抗体结合，形成抗体-抗原-酶标抗体复合物，经过彻底洗涤后加底 ...

小鼠卵巢癌标记物CA125酶联免疫分析（ELISA）试剂盒使用说明书本试剂仅供研究使用目的：本试剂盒用于测定小鼠血清，血浆及相关液体样本中卵巢癌标记物CA125的含量。实验原理：本试剂盒应用双抗体夹心法测定标本中小鼠卵巢癌标记物CA125水平。用纯化的小鼠卵巢癌标记物CA125抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入卵巢癌标记物CA125，再与HRP标记的卵巢癌标记物 ...

小鼠破伤风(Tetanus)酶联免疫分析（ELISA）试剂盒使用说明书本试剂仅供研究使用目的：本试剂盒用于检测小鼠血清，血浆及相关液体样本中破伤风(Tetanus)水平实验原理：本试剂盒采用双抗体夹心酶联免疫法（ELISA）测定标本中小鼠破伤风(Tetanus)。用纯化的小鼠破伤风(Tetanus)抗体包被微孔板，制成固相抗体，可与样品中破伤风(Tetanus)相结合，经洗涤除 ...