Tissue Total DNA Purification Kit (Unlabel)

Transcription

1 Tissue Total DNA Purification Kit (Unlabel) Catalog Number U assays Version: 10 Intended for research use only

2 Table of Contents Introduction... 3 Background... 3 Principle of the Assay... 3 General Information... 4 Materials Supplied... 4 Storage Instruction... 4 Materials Required but Not Supplied... 4 Precautions for Use... 4 Assay Protocol... 5 Sample Preparation... 5 Assay Procedure... 5 Program... 6 Auto Plate Content... 6 Resources... 7 Plate Layout... 7 U / 7

3 Introduction Background Tissue Total DNA Purification Kit (Pre-filled) is dedicated to the isolation of DNA from tissues that are difficult to be lysed. Samples need to be first treated with proteinase K, followed by adding samples onto Auto Plate/Tube. The protocol dramatically reduces experimental time and enhances consistency and reproductivity of DNA isolation and is suitable for laboratories with large volume of samples. Principle of the Assay The silicon dioxide layer coated on the magnetic beads can absorb negative charged molecular in order to purity nucleic acid from samples. Sample Types: 50 ~ 100 mg tissue samples U / 7

4 General Information Materials Supplied List of component Item Description Amount Auto Plate 96 well plate with reagent buffers. 6 plates Strip 8-channel strip 12 strips Storage Instruction Components under room temperature (15~35 C) can be stored until the expiration date labeled on the box. Repeating of freezing and thawing may cause the activity decay of Proteinase K. Materials Required but Not Supplied Proteinase K: Store at -20 C. Incubation Buffer: Tris buffer, surfactants, ph 8.0. Elution Buffer: Nuclease-Free Water. Precautions for Use Do not use expired kits. When room temperature is below 20 C. Please warm the reagent plate/tube at 42 ~ 60 C for 5 ~ 10 min. Do not shake the reagent vigorously in order to avoid the excess foam formation. Do not expose plate/tube and bottle reagent to air for a long time, to avoid evaporation and changing ph then affecting purification efficiency. All reagents should be transparent and colorless. The existence of colors indicates that the reagent is contaminated. Please replace another plate to continue following procedure. Before use, inspect the completeness of the Auto plate/tube and strips. Please wear a mask and disposable gloves when manipulation. Remove the aluminum foil carefully to avoid splashing of the reagent solution. Please use sterile consumables, and make sure that they are all nuclease free. The procedures should not be changed. Because the reagent buffers contain guanidine salts, it is prohibited from washing with any detergents that contain bleach. All reagents are to avoid contact with the eyes, skin, and clothes. If any contact or splashing has occurred, rinse with abundant amount of water. U / 7

5 Assay Protocol Sample Preparation 1. Add 200 μl incubation buffer and 10 μl Proteinase K into 1.5 ml tube. 2. Put 50~100 mg tissue into 1.5 ml tube and mix well. 3. After incubation at 56 C for 2~4 hours or overnight, centrifuged at 8000 RPM for 1 minute. Assay Procedure 1. Carefully remove the aluminum foil from Auto plate. 2. Use micropipette to load 200 μl lysate into column #1/#7 of Auto Plate. 3. Push Auto Plate completely to the bottom of plate rack. Make sure that the missing corner of Auto Plate faces toward the door panel. 4. Push strips completely to the bottom of strip rack frame. 5. Close the door panel. 6. Select the program L-BNA-PK-AUTO. The program parameters are given in following section. 7. Once the program has ended, the buzzer shall alarm. Take out Auto plate carefully. 8. Use micropipette to transfer the purified nucleic acid from column #6/#12 to a clean tube. 9. Put the used Auto plates and strips into the waste recovery can. U / 7

6 Program Program Name:L-BNA-PK-AUTO Model:SLA-16/32, SLA-E132 Series Step Well Temp( C) Mixing (M) Collect (S) Rod Mixing Speed Volume Pause Vapor (M) ON Medium 800 OFF OFF Medium 800 OFF OFF Low 900 OFF ON Medium 800 OFF ON Medium 900 OFF ON Medium 800 OFF ON Medium 800 OFF ON Medium 800 OFF ON Medium 800 OFF ON Medium 200 OFF NA 1 0 OFF Medium 800 OFF NA 0 0 OFF Medium 0 OFF 0 Auto Plate Content Column Buffer solution Volume 1/7 Lysis Buffer 700 μl 2/8 Wash Buffer μl 3/9 Magnetic Beads 800 μl 4/10 Wash Buffer μl 5/11 Wash Buffer μl 6/12 Elution Buffer 130 μl U / 7

7 Resources Plate Layout U / 7