RPM Yeast Plasmid Isolation Kit For the purification of plasmid DNA from yeast

Transcription

1 RPM Yeast Plasmid Isolation Kit For the purification of plasmid DNA from yeast Revision # L06

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3 RPM Yeast Plasmid Isolation Kit For the purification of plasmid DNA from yeast Application Manual Revision # J10 Product Description # of Preps Cat #. RPM Yeast Plasmid Isolation Kit 12 preps RPM Yeast Plasmid Isolation Kit 100 preps Any Questions? Call Technical Support at (800)

4 TABLE OF CONTENTS 1. Introduction RPM Yeast Plasmid Isolation Kit Introduction Related Products Key Features and Applications RPM Yeast Plasmid Isolation Kit Components Kit Components (Cat # ) Kit Components (Cat # ) Storage of RPM Yeast Plasmid Isolation Kit RPM Yeast Plasmid Isolation Kit Protocol Appendix (i) Product Use Limitation & Warranty visit us on the web at

5 1. Introduction 1.1 RPM Yeast Plasmid Isolation Kit Introduction The RPM Yeast Plasmid Isolation Kit for the isolation of plasmid DNA from yeast is a Rapid Pure Miniprep Kit designed for yeast cells. The plasmid isolated from yeast is suitable for PCR and bacterial transformation. This kit is ideal for screening yeast transformants obtained from two-hybrid screens. 1.2 Related Products Yeast Kits and Accessories Alkali Cation Yeast Transformation Kit Application: High efficiency transformation of library DNA into yeast two-hybrid reporter strains Quantity Cat # 25 preps Yeast Spheroplast Transformation Kit Application: Screening cdna libraries for complementation for construction of YAC libraries Quantity Cat # 25 preps EZ Yeast Transformation Kit Application: High throughput transformation without making competent cells Quantity Cat # 200 preps Whole Cell Yeast PCR Kit Application: High throughput direct PCR amplification of yeast colonies without purifying DNA Quantity Cat # 500 preps Yeast Cell Lysis Preparation Kit Application: Used with GNOME DNA Isolation Kit to isolate genomic DNA from yeast Quantity Cat # 100 preps Any Questions? Call Technical Support at (800)

6 FastDNA Kit Application: Isolation of genomic DNA from yeast or virtually any source Quantity Cat # 100 preps FastRNA Kit, Red Application: Isolation of total RNA from yeast, algae or fungi Quantity Cat # 100 preps FastPROTEIN Red Kit Application: Isolation of protein from yeast Quantity Cat # 50 preps preps Library-in-a-Tube Application: A PCR-ready single-stranded cdna library made from total RNA for single-use PCR reactions. Description Size Cat # Yeast: S. cerevisiae, Stationary Phase 3 x 0.2 ml Yeast: S. cerevisiae, Stationary Phase 3 x 0.5 ml Yeast: S. cerevisiae, Log Phase 3 x 0.2 ml Yeast: S. cerevisiae, Log Phase 3 x 0.5 ml Yeast: S. pombe, Stationary Phase 3 x 0.2 ml Yeast: S. pombe, Stationary Phase 3 x 0.5 ml Yeast: S. pombe, Log Phase 3 x 0.2 ml Yeast: S. pombe, Log Phase 3 x 0.5 ml Yeast: S. albicans, Stationary Phase x 0.2 ml Yeast: S. pombe, Stationary Phase x 0.5 ml Yeast: S. pombe, Log Phase x 0.2 ml Yeast: S. pombe, Log Phase x 0.5 ml Yeast: S. albicans, Stationary Phase x 0.2 ml Yeast: S. pombe, Stationary Phase x 0.5 ml Yeast: S. pombe, Log Phase x 0.2 ml Yeast: S. pombe, Log Phase x 0.5 ml visit us on the web at

7 Replica Plating Apparatus This hardwood replica plating apparatus is made from fallen trees. It can be used with velvet, gauze, or even paper towels. Replica Plating Apparatus (100 mm) Replica Plating Apparatus (150 mm) Velvet Pad for 100 mm plates (6 sq in.) Velvet Pad for 100 mm plates (6 sq in.) Velvet Pad for 150 mm plates (9 sq in.) Velvet Pad for 1050 mm plates (9 sq in.) Yeast Growth Media Standard Formulations YPD (YEPD) Broth Contents/L: 20 g peptone, 10 g yeast extract-y, 20 g dextrose Large Capsules 227 g Capsules 227 g Powder 227 g Pouch, 0.5 L 10 x 0.5 L Pouch, 1.0 L 10 x 1.0 L YPD Agar Contents/L: YPD, 17 g Agar-Y Capsules 227 g Powder 227 g Pouch, 0.5 L 10 x 0.5 L Pouch, 1.0 L 10 x 1.0 L DOB Contents/L: 1.7 g YNB, 5 g Ammonium Sulfate, 20 g Dextrose Powder 227 g Pouch, 0.5 L 10 x 0.5 L Pouch, 1.0 L 10 x 1.0 L Any Questions? Call Technical Support at (800)

8 DOBA Contents/L: DOB, 17 g Agar-Y Powder 227 g Pouch, 0.5 L 10 x 0.5 L Pouch, 1.0 L 10 x 1.0 L YNB Contents/L: Standard Formulation Powder 227 g YNB w/ Ammonium Sulfate Contents/L: YNB, 5 g Ammonium Sulfate Powder 227 g Complete Supplement Mixture (CSM) CSM powder 10 g CSM-HIS powder 10 g CSM-LEU powder 10 g CSM-TRP powder 10 g CSM-URA powder 10 g * Other formulations available Pre-poured Plates YPD Agar 10 plates SDA Medium Complete 10 plates SDA Medium-HIS 10 plates SDA Medium-LEU 10 plates SDA Medium-TRP 10 plates SDA Medium-URA 10 plates * Other formulations available 8 visit us on the web at

9 Yeast Growth Media Additives and Components Agar-Y Capsules 227 g Powder 227 g FOA Powder 1 g AT Powder 50 g Key Features and Applications Purification of plasmid DNA from yeast for PCR and bacterial transformation No phenol/chloroform extraction Ideal for processing putative positive clones from yeast two-hybrid screens No alcohol precipitation 2. RPM Yeast Plasmid Isolation Kit Components 2.1 Catalog # (12 preps) Description Volume Cat # Yeast Lysis Matrix 0.4 g x 12 tubes Alkaline Lysis Solution 4 ml Neutralizing Solution 4 ml GLASSMILK Spin Buffer 5 ml Wash Solution Concentrate* 7 ml SPIN Filters Catch Tubes *Add 7 ml of 100% Ethanol to Wash Solution Concentrate and mix before initial use Any Questions? Call Technical Support at (800)

10 2.2 Catalog # (100 preps) Description Volume Cat # Yeast Lysis Matrix 0.4 g x 100 tubes Alkaline Lysis Solution 32 ml Neutralizing Solution 32 ml GLASSMILK Spin Buffer 32 ml Wash Solution Concentrate 56 ml SPIN Filters Catch Tubes *Add 56 ml of 100% Ethanol to Wash Solution Concentrate and mix before initial use 3. Storage of RPM Yeast Plasmid Isolation Kit The RPM Yeast Plasmid Isolation Kit is shipped and stored at ambient temperature 4. RPM Yeast Plasmid Isolation Kit Protocol 1. Temporarily remove Yeast Lysis Matrix from screw cap tube by pouring into another clean tube or container. Add 1.5 ml of yeast culture to the empty tube and spin for 30 seconds to pellet the cells. Discard the supernatant. 2. Pour the Yeast Lysis Matrix back into the tube, add 250 µl Alkaline Lysis Solution and vortex continuously for 5 minutes or shake for 10 seconds in the FastPrep FP120 Instrument. The presence of detergents in the Lysis Solution will cause the sample to foam. To facilitate the processing of multiple samples, use a multi-tube holder attached to the vortex machine. The FastPrep FP120 Instrument (cat. # ) is a controlled high-speed shaking device available through BIO 101 Systems to extract nucleic acids from virtually any source: cells, tissues, organs, and organisms; it can process up to 12 samples simultaneously. 3. Add 250 µl of Neutralizing Solution; mix by brief vortexing and spin 2 minutes at room temperature. Transfer the supernate to a Spin Filter, avoiding the precipitated debris and Lysis Matrix. 4. Add 250 µl of GLASSMILK Spin Buffer; invert to mix; spin 1 minute and decant the Catch Tube. 5. Add 500 µl of Wash Solution; spin 1 minute and decant the wash. Repeat the wash step; decant the Catch Tube and spin for 1 minute to drive the last of the liquid out of the Spin Filter. Transfer the Filter to a new Catch Tube. 10 visit us on the web at

11 6. Add 100 µl of sterile H 2 O; vortex briefly (at no more than half speed) to resuspend and spin 30 seconds to collect the DNA in the bottom of the Catch Tube; discard the Filter containing used Matrix. Use 5 µl for transformation and PCR analysis. Note: Electroporation is preferred over CaCl 2 -heat-shock treatment when transforming bacterial cells. 5. Appendix (i) Product Use Limitation & Warranty Unless otherwise indicated, this product is for research use only. Purchase of BIO 101 Systems products does not grant rights to reproduce, modify, repackage the products or any derivative thereof to third parties. BIO 101 Systems makes no warranty of any kind, expressed or implied, including merchantability or fitness for any particular purpose, except that the products sold will meet our specifications at the time of delivery. Buyer s exclusive remedy and BIO 101 Systems sole liability hereunder shall be limited to, at our option, product credits, refund of the purchase price of, or the replacement of all material(s) that does not meet our specification. By acceptance of the product, Buyer indemnifies and holds BIO 101 Systems harmless against, and assumes all liability for the consequence of its use or misuse by the Buyer, its employees or others, including, but not limited to, the cost of handling. Said refunds or replacement is conditioned on Buyer notifying BIO 101 Systems within thirty (30) days of receipt of product. Failure of Buyer to give said notice within thirty (30) days shall constitute a waiver by the Buyer of all claims hereunder with respect to said material(s). Any Questions? Call Technical Support at (800)

12 Notes 12 visit us on the web at

13 Notes Any Questions? Call Technical Support at (800)

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