| CAT No | Product | Size | Price |
|---|---|---|---|
| PB72.10-01 | VeriFi® Library Amplification Mix | 50 x 50 μL Reactions | Contact us |
| PB72.10-05 | VeriFi® Library Amplification Mix | 250 x 50 μL Reactions | Contact us |
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Technical Specifications
VeriFi® Library Amplification Mix
| Component | 50 Reactions | 250 Reactions |
|---|---|---|
| 2x VeriFi Library Amplification Mix | 1 x 1.25 mL | 5 x 1.25 mL |
VeriFi® Library Amplification Mix
| Component | 50 Reactions | 250 Reactions |
|---|---|---|
| 2x VeriFi Library Amplification Mix | 1 x 1.25 mL | 5 x 1.25 mL |
Reaction Information
| Reaction Volume | Storage | |||
|---|---|---|---|---|
| 50 μL | Upon receipt, the product should be stored at a temperature between -30 and -20 °C. If stored correctly, the kit will maintain full activity until the specified expiry date. |
Frequently Asked Questions (FAQs)
Can I perform multiplex PCR with VeriFi® Library Amplification Mix?
Yes. For most multiplex experiments, we recommend using VeriFi® Hot Start Polymerase & Mixes (product numbers PB10.45-PB10.47). However, VeriFi® Library Amplification Mix is also well-suited for this purpose, particularly when one or more multiplex targets contain GC-rich regions or have challenging or repetitive sequence regions.
Can I use VeriFi® Library Amplification Mix for bisulfite-treated DNA methylation studies/amplification?
No. Pfu polymerases can only amplify targets containing uracil following transformation, which reduces PCR accuracy.
Can I use VeriFi® Library Amplification Mix for standard PCR instead of library amplification?
Yes, please follow the relevant guidance in the product manual. VeriFi® Library Amplification Mix can be used in standard 3-step or 2-step cycling programs and is particularly suited for amplifying targets with very high or very low GC content.
Can VeriFi® Library Amplification Mix be used in single-cell NGS experiments?
VeriFi® Library Amplification Mix can be used for PCR-based enrichment of any type of DNA library.
Do I need to include a purification step between adaptor ligation and library amplification with VeriFi® Library Amplification Mix?
We recommend including a purification step after adaptor ligation and size selection. This removes any buffer incompatibilities and ensures that unligated adaptors are not amplified during the PCR library enrichment.
I am having difficulty amplifying a target with high GC content, what should I do?
Increase the denaturation temperature to 98-100 °C and increase the extension time.
My amplified library is of low quality, how can I improve it?
Increase denaturation temperature. Optimize annealing temperature and extension time. Optimize primer and template amounts in the reaction. Ensure the starting material is of high quality.
What is the maximum target length I can amplify with VeriFi® Library Amplification Mix?
We have successfully amplified up to a 12 kb fragment containing GC-rich regions. Longer targets can also be successfully tested.
What is the minimum sample amount I can use for successful library amplification with VeriFi® Library Amplification Mix?
Do not use less than 10 ng of template for NGS library amplification. A key factor in obtaining high-quality amplified libraries is limiting the number of PCR cycles used. Using higher cycle numbers will result in a greater risk of errors and bias caused by PCR.
What NGS sequencing platforms can I use VeriFi® Library Amplification Mix with?
VeriFi® Library Amplification Mix has been extensively tested and validated on Illumina sequencing platforms with Illumina P5 and P7 primers. The mix should also work well in amplifying libraries with different adapter sequences intended for other sequencing platforms (e.g., Pac Bio and Oxford Nanopore Technologies), but this should be verified.
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