For 250 preps: 130 ml Buffer GB, 50 ml DNA Wash Buffer concentrate (for 250 ml 1x) and 20 ml Elution Buffer. 

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For 250 preps: 125 ml Buffer PB, 50 ml DNA Wash Buffer concentrate (for 1x250 ml) and 20 ml Elution Buffer. 

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The buffer set includes: 250 ml A1  250 ml A2  300 ml A3  900 ul RNase A Good for 24 Maxipreps, 48 Midiprep-II preps and 96 Midipreps

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 For 250 preps: 80 ml A1, 80 ml A2, 110 ml N1, 50 ml washing buffer concentrate (for 250 ml washing buffer), 20 ml Elution buffer, 280 µl RNase A.

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Includes Buffer LY, RNA Washing Buffer and Elution Buffer.

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Includes Buffer LY, RNA Washing Buffer and Elution Buffer.

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Grade: AR CAS No.: 10035-04-8 Synonym: Calcium Chloride dihydrate Molecular Fomula: CaCl2.2H2O Molecular Weight: 147.01 g/mol Storage condition: Room temperature Analytical Specifications Specification Result Appearance White powder or block Conform Content ≥98.0% 99.4% Alkalinity Comply with the standard Conforms Iron ≤0.002% 0.001% Magnesium and alkali metals ≤0.3% 0.2% Safety Data Sheet MSDS AR Grade- The standard Mallinckrodt grade of analytical reagents; suitable for laboratory and general use. Analytical Research Grade chemicals are a very high grade of chemical suitable for the specialist analytical testing carried out in laboratories where a high degree of accuracy is required.

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Grade: AR CAS No.: 10035-04-8 Synonym: Calcium Chloride dihydrate Molecular Fomula: CaCl2.2H2O Molecular Weight: 147.01 g/mol Storage condition: Room temperature Analytical Specifications Specification Result Appearance White powder or block Conform Content ≥98.0% 99.4% Alkalinity Comply with the standard Conforms Iron ≤0.002% 0.001% Magnesium and alkali metals ≤0.3% 0.2% Safety Data Sheet MSDS AR Grade- The standard Mallinckrodt grade of analytical reagents; suitable for laboratory and general use. Analytical Research Grade chemicals are a very high grade of chemical suitable for the specialist analytical testing carried out in laboratories where a high degree of accuracy is required.

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Grade: AR CAS No.: 10035-04-8 Synonym: Calcium Chloride dihydrate Molecular Fomula: CaCl2.2H2O Molecular Weight: 147.01 g/mol Storage condition: Room temperature Analytical Specifications Specification Result Appearance White powder or block Conform Content ≥98.0% 99.4% Alkalinity Comply with the standard Conforms Iron ≤0.002% 0.001% Magnesium and alkali metals ≤0.3% 0.2% Safety Data Sheet MSDS AR Grade- The standard Mallinckrodt grade of analytical reagents; suitable for laboratory and general use. Analytical Research Grade chemicals are a very high grade of chemical suitable for the specialist analytical testing carried out in laboratories where a high degree of accuracy is required.

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Intended Use The Master Mix is used for real-time qualitative and quantitative RT-PCR amplifications with SYBR Green dye for extraction-free RNA samples. The master mix is a premixed, 2X concentrated solution that has all the components except for gene-specific primers. Kit Features The master mix contains extra more redundant amounts of the RTase and DNA polymerase, specifically designed to overcome endogenous inhibition in extraction-free RNA samples. For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which is a thermophilic type A polymerase, with optimal temperatures of 60-62°C. The RTase is easily heat-inactivated at ≥90°C for 1min. The RTase efficiently synthesizes a complementary DNA strand on RNA template from a gene-specific primer, ≤1 unit per 20μL of reaction. The RTase reversely-transcribes single digit copies of target RNA molecules consistently. The kit also contains Taq-Fast DNA polymerase which extends more than 300 bases with short cycling program. The concentrations of the primers are variable depending on assay designs and thermocycling protocols (Table 1). The preferable PCR product size is ≤150bp. The kit has three formulations of No ROX, Low ROX or High ROX concentrations for your choice.Kit Contents 2X Master Mix (2x1mL/5x1mL/10mL for 200/500/1000 reactions x 20μL) and an instruction for use. The kit also contains Lysis Buffer (2x10mL/1x50mL/1x100mL). Transportation and Storage The kit can be transported at below 4°C for up to 3 days. The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a week. Protocol for extraction-free RNA from cultured cells Wash 10,000-20,000 adherent cells in a well or suspension cells in a medium with 1X PBS buffer. Add 50-100ul Lysis Buffer to the well or to the collected cells, and leave for 5min. Break down the cells through either sonicator, or vigorous vortex for 1 min. Heat at 95ºC for 10min and then cool down to 4°C. Up to 4μl of lysed RNA sample can be added to a 20ul final RT-PCR volume.Note: the recovery rate of the extraction-free RNA is heavily affected by how completely the cells are broken down. Setup Reaction and Thermocycling Thaw 1-Step 2X RT-PCR Master Mix and other reaction components at room temperature, mix each component, centrifuge and then place on ice. Set up reactions (Table 1). Seal tubes with flat, optically transparent caps or seal plate with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-900nM RNA Templatesb Variable Variable As low as single digit copies of target RNA to ≤1μg total RNA H2O To 20μL To 10μL Notes: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity. Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 60°C 10min 1 II 95°C 1min 1 III 95°C 10sec 35-40 III 60°C, signal acquisition 60sec 35-40 IV 60°C to 95°C Various 1 Table 3. Three-Step Thermocycling Protocol Stage Temperature Period Number of Cycles I 60°C 10min 1 II 95°C 1min 1 III 95°C 10sec 35-40 III 60°C 30 sec 35-40 III 68-72°C, signal acquisition 30 sec 35-40 IV 68°C to 95°C Various 1 Notes: The three-step thermocycling protocol in Table 3 increases overall polymerase activity by 50%, a more effective protocol than Table 2. The primer concentration used in Tables 2 and 3 is typically 0.15-0.2uM.   Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, Opticon, Roche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye  Precautions If you order a “No ROX” master mix but you have an Applied Biosystems or Thermo Fisher instrument, please turn off ROX passive reference dye button when setup assays.

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Intended Use The Master Mix is used for real-time qualitative and quantitative RT-PCR amplifications with SYBR Green dye for extraction-free RNA samples. The master mix is a premixed, 2X concentrated solution that has all the components except for gene-specific primers. Kit Features The master mix contains extra more redundant amounts of the RTase and DNA polymerase, specifically designed to overcome endogenous inhibition in extraction-free RNA samples. For the reverse transcription step, this kit uses a highly efficient Thermophilic Reverse Transcriptase (US patent pending), which is a thermophilic type A polymerase, with optimal temperatures of 60-62°C. The RTase is easily heat-inactivated at ≥90°C for 1min. The RTase efficiently synthesizes a complementary DNA strand on RNA template from a gene-specific primer, ≤1 unit per 20μL of reaction. The RTase reversely-transcribes single digit copies of target RNA molecules consistently. The kit also contains Taq-Fast DNA polymerase which extends more than 300 bases with short cycling program. The concentrations of the primers are variable depending on assay designs and thermocycling protocols (Table 1). The preferable PCR product size is ≤150bp. The kit has three formulations of No ROX, Low ROX or High ROX concentrations for your choice.Kit Contents 2X Master Mix (2x1mL/5x1mL/10mL for 200/500/1000 reactions x 20μL) and an instruction for use. The kit also contains Lysis Buffer (2x10mL/1x50mL/1x100mL). Transportation and Storage The kit can be transported at below 4°C for up to 3 days. The kit should be kept stable in the dark at -20°C for ≤24 months with ≤10 times of freeze-thaw cycles. The kit can be stored at 4°C for a week. Protocol for extraction-free RNA from cultured cells Wash 10,000-20,000 adherent cells in a well or suspension cells in a medium with 1X PBS buffer. Add 50-100ul Lysis Buffer to the well or to the collected cells, and leave for 5min. Break down the cells through either sonicator, or vigorous vortex for 1 min. Heat at 95ºC for 10min and then cool down to 4°C. Up to 4μl of lysed RNA sample can be added to a 20ul final RT-PCR volume.Note: the recovery rate of the extraction-free RNA is heavily affected by how completely the cells are broken down. Setup Reaction and Thermocycling Thaw 1-Step 2X RT-PCR Master Mix and other reaction components at room temperature, mix each component, centrifuge and then place on ice. Set up reactions (Table 1). Seal tubes with flat, optically transparent caps or seal plate with optically transparent film. Mix and then briefly centrifuge the tubes or plate. Program PCR instrument with indicated thermo-cycling protocol. Load PCR tubes or plate and start to run. Perform data analysis according to the PCR instrument instructions. Table 1. Setting up a 20μl or 10μl Reaction Component Volume per 20μl Volume per 10μl Final Concentration 2x Mastermix 10μL 5μL 1X Primersa Variable Variable Each 150-900nM RNA Templatesb Variable Variable As low as single digit copies of target RNA to ≤1μg total RNA H2O To 20μL To 10μL Notes: Each primer’s Tm should be designed ≥60°C, preferably between 62°C to 65°C, using primer3 software for high efficiency and specificity. RNA templates should be extracted by a qualified silica-based kit and eluted with low EDTA TE buffer (10mM Tris-HCl, 0.1mM EDTA, pH 8.0-8.3). Table 2. Standard Thermocycling Protocol Stage Temperature Period Number of Cycles I 60°C 10min 1 II 95°C 1min 1 III 95°C 10sec 35-40 III 60°C, signal acquisition 60sec 35-40 IV 60°C to 95°C Various 1 Table 3. Three-Step Thermocycling Protocol Stage Temperature Period Number of Cycles I 60°C 10min 1 II 95°C 1min 1 III 95°C 10sec 35-40 III 60°C 30 sec 35-40 III 68-72°C, signal acquisition 30 sec 35-40 IV 68°C to 95°C Various 1 Notes: The three-step thermocycling protocol in Table 3 increases overall polymerase activity by 50%, a more effective protocol than Table 2. The primer concentration used in Tables 2 and 3 is typically 0.15-0.2uM. Table 4. Compatible Instruments qPCR Instrument ROX required by instrument Passive dye setup Bio-Rad® iQ™5, CFX96, CFX384, Opticon, Roche Lightcycler®Qiagen Rotor-Gene™Eppendorf Mastercycler®Cepheid® SmartCycler® Not recommended Not necessary Applied Biosystems® 7500, 7500 Fast, QuantStudio™, ViiA7™, Agilent Mx™ Low ROX(50nM final concentration) Turn on ROX passive reference dye  Applied Biosystems® 5700, 7000, 7300, 7700, 7900, 7900HT, 7900HT Fast, StepOne™, StepOnePlus™ High ROX(500nM final concentration) Turn on ROX passive reference dye  Precautions If you order a “No ROX” master mix but you have an Applied Biosystems or Thermo Fisher instrument, please turn off ROX passive reference dye button when setup assays.

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Used with dishes and cell culture clusters for quick removal of cell layers Blade Length: 2.0cm; Handle Length: 16cm Gamma radiation sterilized and individually wrapped  Disposable lifters quickly remove cell layers The one-piece design and chiseled blade provide improved control while minimizing damage during the cell removal process  Non-pyrogenic  Polyethylene

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