Regulatory requirements vary by product. Please see product-specific literature to determine product use.

Product FAQ

Can I autoclave agarose to prepare agarose gels?

Answer

No. Autoclaving at temperatures that are too high or pressures that are too high can hydrolyze the agarose polymer and weaken it.

Answer Id: E2921

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

Why do solutions containing SDS form precipitates?

Answer

SDS is a detergent used to denature proteins leading to disruption of cell membranes and dissociation of nucleic acid-protein complexes in DNA and RNA extractions. It tends to come out of solution at concentrations of 10% or greater, especially at low temperatures or in high-salt buffers. SDS can be resolubilized by slowly heating the solution to 65°C with slight agitation.

Answer Id: E2924

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What should the running conditions be for the TBE gels (voltage, current, run time, etc.)?

Answer

Voltage: 200 V constant*
Approximate current at start: 10-18 mA/gel
Approximate current at end: 4-6 mA/gel
Run time: Approximately 30-90 minutes, dependent on gel percentage. The run is complete when the bromophenol blue (darker) tracking dye reaches the bottom of the gel.

* Voltages up to 250 V may be used to reduce run time.

Answer Id: E7995

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

My Xeno control assay failed. What are some possible causes?

Answer

If your Xeno control fails, you could possibly have inhibitors in your samples. You can test this by running the Xeno control directly in a qPCR reaction. You can also run a mock extraction control where you spike the Xeno control into a buffer sample and then perform the nucleic acid extraction procedure you used for your samples.

Answer Id: E16422

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

How can I tell if my RT-PCR product is RNA specific?

Answer

Include a control reaction where the RNA has not been incubated with reverse transcriptase to test for specificity. If this RNA gives a PCR product, it is most likely generated from genomic DNA contamination. Alternatively, a primer set spanning two different exons can be designed such that the PCR product from the cDNA would be of a different size compared to a product generated from genomic DNA. Primers may also be designed to span an exon/exon junctions. These primers are not likely to amplify from genomic DNA templates. For DNase treatment of RNA, we recommend using Amplification-Grade DNase I, Cat. No. 18068-015, or an equivalent product.

Answer Id: E2926

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

Does the Platinum Multiplex PCR master mix leave A overhangs?

Answer

Yes, the Platinum Multiplex PCR master mix contains Platinum Taq DNA Polymerase that leaves A overhangs on PCR products.

Answer Id: E7626

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

Can I stain my TBE gel or my TBE-urea gel? How?

Answer

Yes, for ethidium bromide staining, soak the gel in a 2 µg/mL solution of ethidium bromide in ultrapure water for 20 minutes. Destain by rinsing with three successive 10-minute rinses of ultrapure water. Visualize bands under UV light

Answer Id: E7996

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

I need support regarding my freezer, refrigerator, incubator, CO2 incubator, centrifuge, oven, furnace, bio-safety cabinet or stirrer. Where can I find help?

Answer

For laboratory equipment products, please go to https://kb.unitylabservices.com

Answer Id: E16426

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

Which is better to use, T4 or E. coli DNA ligase?

Answer

It depends on your application. For ligation of dsDNA fragments with cohesive ends, either enzyme can be used. E. coli DNA ligase requires the presence of beta-NAD, while T4 DNA ligase requires ATP. However, only T4 DNA ligase can join blunt-ended DNA fragments - E. coli ligase is unable to join such fragments.

E. coli DNA ligase is generally used to eliminate nicks during second-strand cDNA synthesis. T4 DNA ligase should not be substituted for E. coli DNA ligase in second-strand synthesis because of its capability for blunt end ligation of the ds cDNA fragments, which could result in formation of chimeric inserts.

Answer Id: E2949

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What is the highest temperature that SuperScript III , SuperScript II, MMLV, or ThermoScript can be used?

Answer

The optimal temperature for SuperScript III RT is 50 degrees C, and can be used up to 55 degrees C. For some qRT-PCR reactions where gene-specific primers are used, you can do the RT reaction at 60 degrees C. The optimal temperature for SuperScript II RT is 42 degrees C, and can be used up to 50 degrees C. Optimal temperature for MMLV is 42 degrees C. ThermoScript RT shows optimal activity at 60 degrees C, and can be used at temperatures as high as 70 degrees C (for amplicons expected to be 1 kb or less). For PCR products expected to be greater than 1 kb, a maximum first strand synthesis temperature of 60-65 degrees C is suggested. Be sure your first-strand primer anneals at the high temperature, especially when gene-specific primers are used for high-temperature stable reverse transcriptases. We recommend oligo (dT)20 for cDNA synthesis when using an oligo (dT) primer for first-strand synthesis with these enzymes.

Answer Id: E2928

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

I am interested in generating cDNA from total RNA. What is the difference between SuperScript III Reverse Transcriptase and SuperScript III First Strand Synthesis System for RT-PCR?

Answer

SuperScript III Reverse Transcriptase (Cat. Nos. 18080093, 18080044, 18080085) contains the stand-alone enzyme and a vial each of 5X first-strand buffer and 100 mM DTT.

SuperScript III First Strand Synthesis System for RT-PCR is a complete kit that provides the SuperScript III Reverse Transcriptase and all the other components required for synthesis of first-strand cDNA from total or poly(A)- RNA. It includes:
- Superscript III Reverse Transcriptase
- Oligo (dT)20 Primer
- Random hexamers
- 10X RT buffer
- 25 mM MgCl2
- 0.1 M DTT
- 10 mM dNTP Mix
- RNAseOUT Recombinant Ribonuclease Inhibitor
- E. coli RNAse H
- DEPC-treated water
- Total HeLa RNA control
- Sense control primer
- Anti-sense control primer
Note: The kit does not include the PCR amplification enzyme.

Answer Id: E7627

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What is the concentration of EDTA in the TBE gels? How about glycerol?

Answer

The EDTA concentration in our TBE gels is 0.06% (w/v). The 20% TBE gels contain 4% glycerol for maximal resolution. All other TBE gels contain 0.8% glycerol in a layer that represents the bottom 9% of the gel. There is no glycerol in the rest of the gel.

Answer Id: E7997

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What is the maximum culture volume I can put in a flask in the Expi293 Expression System?

Answer

This depends on the flask, but a general rule of thumb is to use one quarter the volume of the flask.

Answer Id: E9238

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

I need support regarding my AA / ICP / DCP / DC Arc / EDS / XRF / XRD / OE and Combustion technique products. Where can I find help?

Answer

Please go to https://kb.unitylabservices.com/?cid=12825&c=13142&cpc=iCia805H4XUv1Ub1aS0kPQGY1OLg7MkTjK for support on our elemental analysis products.

Answer Id: E16427

Was this answer helpful?

Yes
No
Thank you for your response

Product FAQ

What are the recommended conditions for cohesive-end ligations?

Answer

Generally, ligations are done in a 20 µL volume. Use a total of 10 to 100 ng of DNA per reaction with an insert to vector ratio of 3:1. Add 0.1 units (Weiss) ligase to the reaction. Incubate at room temperature for 30-60 minutes.

Optimal ligation may occur at other ratios (e.g. 1:5, 1:10). If possible, assemble several ligation reactions of varying insert to vector ratios in order to reveal the optimal ligation conditions.

Thermo Fisher Scientific offers T4 DNA ligase at two concentrations: 1 U/µL (Cat. No. 15224-017) and 5 U/µL (Cat. No. 15224-041). When performing blunt or TA cloning ligations, the higher concentration of ligase is generally preferred since ligating a blunt or single base overhang requires more enzyme.

Answer Id: E2950

Was this answer helpful?

Yes
No
Thank you for your response