sgRNA Synthesis Kit, spCas9
Cat. No.
G520
Unit
10 reactions
Price
$149.00
$99.00
| Cat. No. | G520 | ||||||||||||||||
| Name | sgRNA Synthesis Kit, spCas9 | ||||||||||||||||
| Unit | 10 reactions | ||||||||||||||||
| Category | Molecular Biology Enzymes and Kits | ||||||||||||||||
| Description |
The sgRNA Synthesis Kit, spCas9 is a complete kit designed for high yield in vitro RNA transcription utilizing T7 RNA Polymerase. This kit includes all reagents required to generate a DNA template and perform RNA transcription with subsequent DNA removal. Resulting products can be used directly for downstream CRISPR/Cas9 studies and/or RNA experiments. This kit contains reagents sufficient for 10 reactions generating up to 100 µg RNA per reaction.
Additional Materials Required (not included)
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| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. G520 |
Datasheet
Search CoA here
| How should I store this product? | |
|
This product should be stored at -20°C to maintain its activity and stability. Ensure that the enzyme is kept in a frozen state until ready to use.
|
| What is the expected yield of sgRNA per reaction? | |
|
Each reaction can generate up to 100 µg of sgRNA, depending on reaction conditions and template quality.
|
| Can this kit be used for generating sgRNA for other Cas proteins? | |
|
This kit is specifically optimized for spCas9 sgRNA synthesis. While the general process may work for other Cas proteins, modifications to the scaffold sequence may be necessary.
|
| What should I do if I see low yield after in vitro transcription? | |
|
Ensure that:
|
| What kind of DNA templates can be used for sgRNA synthesis? | |
|
Synthetic DNA oligonucleotides, PCR products, or linearized plasmid DNA containing a double-stranded T7 promoter region upstream of the template sequence can be used. SP6 and T3 promoters are not compatible with this kit. |
| How do I check if my PCR amplification was successful? | |
|
Run 5 μl of PCR product on a 2% agarose gel. You should see a 130 bp amplicon. If no bands appear, check primer design and PCR conditions.
|
| Can I store the synthesized sgRNA for later use? | |
|
Yes, the synthesized sgRNA should be stored at -80°C in small aliquots to prevent degradation. Avoid repeated freeze-thaw cycles.
|
| Do I need to purify the sgRNA after transcription? | |
|
For most CRISPR applications, the RNA can be used directly. However, if higher purity is required, you may use LiCl precipitation, phenol-chloroform extraction, or column-based RNA purification kits.
|
| What RNase precautions should I take when handling sgRNA? | |
|
| Can I use the synthesized sgRNA for in vivo applications? | |
|
Yes, but additional RNA purification and quality control (e.g., gel electrophoresis, spectrophotometry, or Bioanalyzer analysis) may be required for in vivo or sensitive applications.
|
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