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PCR/qPCR
MIDSCI can supply almost every needed product for PCR/qPCR: tubes, plates, strips, caps, taq, Mastermixes, dNPTs, dye and probe for qPCR, sealing film and much more. If you are looking for any of these things, we can match almost all needs to all cyclers. If you don’t find it easily contact us at tech@midsci.com and we will find it for you.
96 or 384 we have it. Non-Skirted, semi-skirted or full skirted, we have it. 0.1, 0.2, or 0.5 mL we have it. A1, A12, H12, A24, P24 cuts, we have them. Sybr© Green or probe alternative, we have it.
Pro Tip: Colored plastics do not have any impact on DNA amplification. But when setting up qPCR it is often recommended to use white plastics. This will limit or prevent fluorescence refraction. Light refraction can minimize sensitivity and consistency.
Pro Tip 2: If you are seeing inconsistencies in PCR/qPCR try PR1MA Pipette Tips. This will help optimize and reduce variance in the reactions. Simple test, see the residual liquid left in tips? This means some of the taq, dNTPs, sample, etc is left in the tip. PR1MA tips help reduce that significantly and improve your consistency.
Pro Tip 3: Seeing evaporation in the outer wells of a PCR/qPCR plate? Sometimes it’s the plate, sometimes it the seal, and sometimes it’s the cycler. Call us we have solutions.
- Excellent all-purpose amplification enzyme
- Thermostable recombinant DNA polymerase from Thermus aquaticus
- Exhibits very high activity in primer extension
- Has both a 5' to 3' DNA polymerase and a 5' to 3' exonuclease activity
- Does not have 3' to 5' exonuclease activity-no proofreading ability
- Leaves an A-overhang, which makes the enzyme ideal for TA cloning
- Includes MgCl² in buffer
Need great (q)PCR Regents at a great price? Try PR1MA! Click here to order.
Store at -20°C. For in-vitro laboratory use only
General Description
Bullseye Taq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. The enzyme is isolated from Thermus aquaticus and has a molecular weight of approximately 94 kDa.
Bullseye Taq DNA Polymerase has both a 5'®3' DNA polymerase and a 5'®3' exonuclease activity. The enzyme lacks a 3'®5' exonuclease activity (no proofreading ability). Taq DNA Polymerase leaves an A¢ overhang, which makes the enzyme ideal for TA cloning.
The 10X Reaction Buffer provided does not contain Mg+2. 25 mM MgCl2 is supplied separately.
10X Mg++ Free Standard Buffer
100 mM Tris-HCl pH 8.3, 500 mM KCl.
Unit Definition
One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions.
Storage and Dilution Buffer
Enzyme is supplied in 20 mM Tris-HCl pH 8.3, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20, 0.5%NP40, 50% glycerol.
Quality Control
Each lot of Taq DNA Polymerase is tested for contaminating activities, with no trace of endonuclease activity, nicking activity, exonuclease activity or priming activity.
Suggested Protocol using Taq DNA Polymerase
This protocol serves as a guideline only. Optimal reaction conditions may vary and must be individually determined.
n In some applications, MgCl2 is needed for the best results. For this reason, 25 mM MgCl2 is included with the kit.
1. Thaw 10X Mg2+ Free Standard Buffer, dNTP mix, primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.
2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA.
Table 1. Reaction components (master mix & template DNA)
Component |
Vol./reaction |
Final Conc. |
10X Mg2+ Free Buffer |
5 µl |
1X |
MgCl2, 25 mM |
1- 9 µl |
0.5 4.5 mM |
dNTP mix (12.5 mM of each) |
0.8 µl |
0.2 mM of each dNTP |
Primer A |
Variable |
0.11.0 µM |
Primer B |
Variable |
0.11.0 µM |
Taq DNA Polymerase |
Variable |
1-5 units |
Template DNA |
Variable |
Variable |
Distilled Water |
Variable |
- - - - |
TOTAL volume |
50 µl |
- - - - |
Table 2. MgCl2 concentration in a 50 mL reaction
Final MgCl2 conc. in reaction (mM) |
0.5 |
1.0 |
1.5 |
2.0 |
2.5 |
3.0 |
3.5 |
4.0 |
4.5 |
Volume of 25 mM MgCl2 per rxn (µl): |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
9 |
Table 2 provides the volume of 25 mM MgCl2 to add to the master mix if a certain MgCl2 concentration is required.
3. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently, e.g., by pipetting the master mix up and down a few times.
4. Add template DNA to the individual tubes containing the master mix.
5. Program the thermal cycler according to the manufacturers instructions. For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.
6. Place the tubes in the thermal cycler and start the reaction.
2x Master Mix Kit (1.5 mM MgCl2)
Bullseye Taq DNA Polymerase Mix is a ready-to-use 2x reaction mix. Simply add primers, template, and water to successfully carry out primer extensions and other molecular biology applications.
Bullseye Taq polymerase, the NH4+ buffer system, dNTPs, and magnesium chloride are conveniently present in the Taq DNA Polymerase Mix. (Inert Red Dye is present in BE180303 only)
Bullseye Taq DNA Polymerase Mix offers several advantages. Set up time is significantly reduced. The chance of contaminating component stocks is eliminated. Reduction of reagent handling steps leads to better reproducibility. Standard tests can be set up with the confidence that results will be consistent every time.
Composition of 2x Taq Master Mix
150 mM Tris-HCl pH 8.5, 40 mM (NH4)2S04, 3 mM MgCl2, 0.2%
Tween 20Ò
4 mM dNTPs
2 units/µL AS ONE Taq polymerase
Inert Red Dye & Stabilizer (BE180303 only)
R-Taq DNA Polymerase 5 units/µL
Cat. No. | Units | 10X Ammonium Buffer (MgCl2 15mM) | MgCl2 25 mM |
BE200303 | 500 | 1.5 mL | 1.5 mL |
BE200304 | 1,000 | 2x 1.5 mL | 2x 1.5 mL |
BE200306 | 2,500 | 4x 1.5 mL | 4x 1.5 mL |
Store at -20°C. For in-vitro laboratory use only
General Description
Bullseye R-Taq DNA Polymerase is a thermostable recombinant DNA polymerase, which exhibits very high activity in primer extension and other molecular biology applications. R-Taq contains a red dye which provides easy and quick identification of reactions to which enzyme was added and allows confirmation of complete mixing. The inert dye has no effect on downstream processes. R-Taq is added directly to the reaction mix and is used in the same manner as standard Taq DNA Polymerase.
Bullseye R-Taq DNA Polymerase has both a 5'®3' DNA polymerase and a 5'®3' exonuclease activity. The enzyme lacks a 3'®5' exonuclease activity. R-Taq DNA Polymerase leaves an A-overhang, which makes the enzyme ideal for TA cloning.
- High performance thermostable DNA polymerase
- Red dye identifies tubes which contain enzyme and confirms complete mixing of reagents
- Leaves an A-overhang
Unit Definition
One unit is defined as the amount that incorporates 10 nmoles of dNTPs into acid-precipitable form in 30 minutes at 72°C under standard assay conditions.
Storage Buffer
Enzyme is supplied in 20 mM Tris-HCl pH 8.3, 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, inert dye, 0.5 % Tweenà 20, 0.5% NP40, 50% glycerol.
Component | Vol./reaction | Final Conc. |
10X Ammonium Buffer | 5 µL | 1X |
dNTP mix (12.5 mM each) | 0.8 µL | 0.2 mM each dNTP |
Primer A | Variable | 0.1-0.5 µM |
Primer B | Variable | 0.1-0.5 µM |
R-Taq DNA Pol | 1 mL | 5 units/reaction |
Distilled Water | Variable | - - - - |
Template DNA | Variable | 0.1-0.5 µg/reaction |
TOTAL volume | 50 µL | - - - - |
Table 2. MgCl2 concentration
3. Mix the master mix thoroughly and dispense appropriate volumes into reaction tubes. Mix gently (e.g., by pipetting) the master mix up and down a few times.
4. Add template DNA (0.1-0.5 mg/reaction) to the individual tubes containing the master mix.
5. Program the thermal cycler according to the manufacturer's instructions.
For maximum yield and specificity, temperatures and cycling times should be optimized for each new template target or primer pair.
6. Place the tubes in the thermal cycler and start the reaction.
7. After primer extension, load 5-10 mL of a 50 mL reaction directly on an agarose gel for analysis.
Tween 20 is a registered trademark of ICI Americas, Inc.
Final MgCl2 conc. in reaction (mM) | 1.5 | 2.0 | 2.5 | 3.0 | 3.5 | 4.0 | 4.5 |
Additional volume of 25 mM MgCl2 per reaction (µL): | 0 | 1 | 2 | 3 | 4 | 5 | 6 |
10X Ammonium Reaction Buffer
Tris-HCl pH 8.5, (NH4)2S04, 15 mM MgCl2, 1% Tween 20
Quality Control
Endonuclease, exonuclease and priming activities are not detected after 3 hours incubation of 1 mg of pUC19 plasmid DNA and 0.5 µg EcoR I digested lambda phage DNA at 72°C in the presence of 40 units of R-Taq DNA Polymerase.
Suggested Protocol using R-Taq Polymerase
This protocol serves as a guideline. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be individually determined.
1. Thaw 10X Ammonium Buffer, dNTP mix, and primer solutions. It is important to mix the solutions completely before use to avoid localized concentrations of salts.
2. Prepare a master mix according to Table 1. The master mix typically contains all the components needed for extension except the template DNA.
The optimal MgCl2 concentration should be determined empirically but, in most cases a concentration of 1.5 mM, as provided in the 1X Ammonium Buffer, produces satisfactory results. Table 2 provides the volume of 25mM MgCl2 to add to the master mix if a higher MgCl2 concentration is required.
Table 1. Reaction components (master mix & template DNA)
PCR SILICONE SEALING MAT - 96
- Made of research-grade silicone
- Scored wells for sample access
- Perforations re-seal after access
- Resists coring and tearing
- Eliminates cross-talk between wells
- Also works as a compression mat
- Suitable for standard PCR thermocycling
- Mats can be reused
- For use with 96-well PCR plates
- Printed alpha numeric grid for easy reference
- Certified to be free of RNase, DNase, DNA, PCR inhibitors, and Pyrogen contamination.
Works as a plate seal (to eliminate cross contamination) and compression mat all-in-one!
Our compression molded 96-round plug, silicone sealing mats for PCR plates minimize sample evaporation in a wide range of temperatures. These mats are chemically resistant and reusable; containing no adhesives or extractables to contaminate samples. Made of research-grade silicone, they can be autoclaved, and following a 10% bleach wash and ethanol rinse protocol may be reused.
Wells are scored (cut 80% of the way through, but not all of the way) to allow for sample access via pipettor, syringe, or probe. These scored perforations will seal themselves, after penetration, ensuring an airtight closure to the well.
These Rotocycler strips are perfectly designed to perform on Rotor-Gene instruments. Tube strips are packaged separately from cap strips. The frosted extensions on caps not only make them more efficient and secure during handling but also offer a convenient area for labeling. For individual use, tube and cap strips can easily be separated and used as individual units.
Click to shop new and improved eXTReme™ 2 Seal Extra Strong Sealing Film
eXTReme™ Seal Sealing Film with Extra Strong Adhesive Minimizing Evaporation Without Residue
eXTReme™ Seal features an extra-strong thick adhesive to minimize evaporation, yet removes easily without leaving residue on the plate.
Features chamfered corners to eliminate plate overhang for robotics applications and tight storage conditions.
- Certified DNase- & RNase- free
- Ideal for use with raised-rim and non-skirted PCR plates
- Available in a larger cut for flat-top semi-skirted and full-skirted PCR plates: eXTReme™ Seal, Large
Application
- PCR
- Robotics
- -20°C to +120°C
Helpful Tips
- Selection of the proper film size will minimize evaporation
- The film should not overhang or ride up the wall of a raised-rim plate
- More complete coverage of skirted flat-top plates will reduce evaporation in the outer wells
- End-tab removal before thermal cycling is highly recommended
ThermalSeal® RT2™ Optically Transparent, Highly Conformable Adhesive, Polyester Sealing Film
Designed for use with standard 96-well plates where rows must be selectively protected or accessed. Available in a smaller cut for raised-rim and non-skirted PCR plates: ThermalSeal® RT2RR
- Certified DNase- & RNase- free
- Ultra-high optical clarity
- Minimizes evaporation and cross-contamination
- Ideal for use with flat-top and hard-shell dual component (e.g. polycarbonate / polypropylene) PCR plates
- Not recommended for removal after thermal cycling
Application
- qPCR
- -40°C to +120°C
Helpful Tips
- Selection of the proper film size will minimize evaporation
- The film should not overhang or ride up the wall of a raised-rim plate
- More complete coverage of skirted flat-top plates will reduce evaporation in the outer wells
- End-tab removal before thermal cycling is highly recommended
myView™ Compact UV Transilluminator
- Uniform Illumination | Four 302 nm Bulbs
- Adjustable UV Safety Cover | Easy Access to Gels
- Large Scratch Resistant Viewing | 16.5 x 13.5 cm
The E3100 Compact UV Transilluminator has a surprisingly small footprint (just over 12 x 8 inches) yet provides industry leading performance. It will accommodate one or two mini gels on the 16.5 x 13.5 cm scratch resistant viewing surface.
The integrated UV blocking cover is made from a polymer that will not fluoresce in UV light. This allows clear viewing of DNA bands through the cover yet offers full UV blocking protection. Two friction hinges on the front edge of the cover allow for full adjustment. Position the cover horizontally for general viewing, tilt it at an angle for gel access and band cutting, or open fully to use the SmartDoc™ Imaging Enclosure (sold separately) for gel imaging with a smartphone.
The transilluminator’s four 6W bulbs evenly illuminate the viewing surface, and the 302 nm wavelength is ideal for Ethidium Bromide, SmartGlow™, SYBR Green®, Gel Green™ and many other common gel stains. Spacing of the UV bulbs, coupled with a reflector system and special diffusing filter glass provides exceptional uniformity across the viewing surface. This allows for quantification of DNA and higher quality imaging when used with a camera system.
Specifications
|
Light source 4 x 6W UV Light Bulbs |
4 x 6W UV Light Bulbs |
|
Safety cover UV blocking polycarbonate |
UV blocking polycarbonate |
|
Exterior dimensions |
10 x 8 x 2.75 in
|
|
Electrical |
12VDC (AC power adapter included) |
|
Output wavelength |
Peak at 302 nm |
|
Viewing surface |
6.5 x 5.3 in. | 16.5 x 13.5 cm |
|
Weight |
0.9 kg | 2 lbs |
|
Warranty |
2 years |
Double sided water cooled vertical system, 10cm x 10cm
- Heavy duty upper buffer chamber with cooling port connections.
- Lower buffer chamber.
- Interlock safety lid with attached leads.
- Casting base.
- Two-pack each of plain & notched glass plates.
- One acrylic blocker plate (used when running only one gel).
- Ttwo 10-well, 0.8mm thick combs
- Four 0.8mm thick side spacers.
- System Accessories
- Choose amoung these accessories to customize each system for your specific needs.
- Alumina cooling plate.
- For use with cooling systems.
- This plate can be sandwiched between the gel and the inner buffer chamber to improve overall temperature uniformity.
- Casting base.
- For quick, efficient and leak-proof casting.
- Multiple gradient caster.
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