Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 207 results for: pcr

Showing: 101 - 150 of 207 results

  1. Protocol - How to Design Primers

    Type
    Protocol
    ...Reaction Plasmid Cloning by PCR Agarose Gel Electrophoresis Primer Design for PCR Oligonucleotide primers ...Tips on designing primers for PCR Protocols...primers are necessary when running a PCR reaction. One needs to design primers that are complementary to the...protocol video below to learn how to design primers for PCR. Reference Page Top Index...

  3. RNA Extraction Without A Kit

    Type
    Blog Post
    ...of a downstream application such as quantitative PCR to assess gene expression. There are several things...

  5. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...reaction. This will allow you to quantify the amount of PCR product that comes from amplification of the original... titer are direct p24 measurements via ELISA and qPCR for viral RNA. Measuring p24 levels in a viral prep...first converted to cDNA and then quantified using qPCR primers targeting specific viral components such...assays depending on the vector backbone. When using qPCR based methods for viral titration consider using...most accurate method of lentiviral titration uses qPCR to measure the number of proviral copies that have...cell line known to contain 1 integrated copy of the qPCR target gene. Developing the proper control line ...

  6. Ligation Independent Cloning

    Type
    Protocol
    ...sterile dH20 to 40 μl Step 4: Amplify Insert by PCR Perform PCR amplification of your insert following the ...ensure a melting temperature between 50-60°C for your PCR primers. Step 2: Linearize Vector In this example...like this: Note: Elute your purified digestion and PCR products with sterile water (instead of TE buffer...important to remove all free nucleotides from your PCR product before proceeding, as they will interfere...Step 5: Create Insert Overhangs Treat your purified PCR product with T4 Pol in the presence of dCTP (not ...

  7. Antibody Production

    Type
    Collection
    ...heavy chain and/or light chain variable regions. The PCR product is purified, sequenced via the Sanger method...

  8. A Look at Addgene's QC Process

    Type
    Blog Post
    ...include places where DNA is being created, such as PCR amplification or bacterial growth. Created with BioRender.com...

  15. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...Watch the Video! Polymerase Chain Reaction (PCR) Basic PCR protocol with tips and FAQ Watch the Video!... digest and ligation Cloning by PCR Generate restriction sites by PCR Modification by Annealed Oligo Cloning...Watch the Video! AAV Titration by qPCR Using SYBR Green Technology Use qPCR to measure the physical titer ...preparation Watch the Video! AAV Titration by ddPCR Use ddPCR to measure the physical titer of your adeno-associated... virus preparation Lentivirus Titration by ddPCR Use ddPCR to measure the physical titer of your lentivirus...

  18. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...Ehrhardt, A., & Baiker, A. (2012). Universal real-time PCR for the detection and quantification of adeno-associated...aliquots of virus. AAV titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing... hours for the qPCR run and data analysis. Refer to this Addgene AAV Titration by qPCR Protocol for more...remember to make one or two small aliquots to use for qPCR titering. This helps avoid multiple freeze/thaws... included. While the physical titer generated by qPCR is useful, I test a few different concentrations...

  24. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ... reference (typically ROX dye), to normalize non-PCR–related fluorescence fluctuations and to minimize...coefficient of correlation) ~ 1.0, E (efficiency of PCR) ~100% (90%–110% range is acceptable) Baseline removal...background signal that is most evident during initial PCR cycles. This background signal must be removed to...Sing A, Ehrhardt A, Baiker A. Universal real-time PCR for the detection and quantification of adeno-associated...Protocols AAV Titration by qPCR Using SYBR Green Technology AAV Titration by qPCR Using SYBR Green Technology...Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min Equipment qPCR instrument Heating plate...genome-containing particles in an AAV prep using qPCR, SYBR Green Technology, and Addgene’s AAV Titration...

  30. CRISPR Plasmids - Plants

    Type
    Collection
    ...BsaI none S. pyogenes Kamoun 52255 pUC119-gRNA U6 PCR template none S. pyogenes Sheen 51295 pRGEB31 rice...

  31. Open Enzyme Collection

    Type
    Collection
    ...this collection include polymerase chain reaction (PCR), cloning, isothermal amplification, reverse transcription...

  34. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...biology 11.1 (2015): 31-37. pCri system The Gomis-Ruth lab designed the pCri system to simplify the protein...the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System Cell Free Protein Expression Kit FREQ Seq...depending upon your specific protein folding needs. The pCri system has recently been used to study a matrix ....8 (2015): 4728-4740. You can find more info on pCri in Mary Gearing’s blog post on the system Easing...Easing the Protein Purification Process with pCri Cell-free protein expression kit Cell-free expression ...

  35. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...Addgene Blog Verify Your Constructs with Colony PCR Read about Tags for Protein Purification Resources... use by the Rinehart lab: pCRT7 NT TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a ...68292 C321.ΔA bacterial strain – Addgene #68306 pCRT7 NT TOPO E17TAG GFP plasmid - Addgene #68295 SupD...

  37. TALEN Plasmids and Kits

    Type
    Collection
    ...somatic cells. Keith Joung Zhang Lab TALE Toolbox PCR/Golden Gate cloning method. Optimized for human expression...

  38. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ..., SacI restriction site and a linker region. The PCR product is then ligated into the MCS/SacI site in...can occur in this system if a small contaminating PCR product formed from primer dimers and containing ...clone. ORFs from an ORFeome Gateway library or any PCR-amplified ORF cloned into pDONR221 can be expressed...

  41. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...and the sequence of gene of interest in separate PCR amplication reactions for recombination in SLIC cloning...the ccdB cassette, which is not copied during the PCR amplification step, so that only vectors with the...or multiply phosphorylated proteins (B40 OTS and pCRT7-GFP). To demonstrate the improvements of the system...

  44. CRISPR References and Information

    Type
    Collection
    ...the tool, you'll need Sanger sequencing reads from PCR amplicons that cover your locus of interest and correspond... PDF, 129 KB Marraffini Bacteria: pCRISPR new spacer cloning pCRISPR PDF, 109 KB Mendenhall and Myers ...

  45. Retrograde AAV viral preps

    Type
    Collection
    ...AAV titering by SYBR green qPCR , confirmation of the transfer plasmid with PCR, purity assessment by silver...

  47. Plan Your Experiment

    Type
    Guide
    ...cell population. The region of interest is PCR amplified, PCR products are denatured-renatured, treated... The PCR product is then run on an agarose gel to determine whether the edit was successful. PCR amplification...run on an agarose gel to identify DNA fragments. PCR and restriction digest (for HDR repaired DSBs): For...novel restriction site. The region of interest is PCR amplified, digested with the appropriate restriction...run on an agarose gel to identify DNA fragments. PCR amplification and gel electrophoresis (for HDR or...deletions or insertions, the region of interest can be PCR amplified using primers that (A) flank the region...and exact sequence of targeted alleles. Involves PCR amplification of targeted region from DNA, subcloning...

  48. Caltech Systemic Capsids

    Type
    Collection
    ...including AAV titering by probe-based droplet digital PCR, in vitro and (when possible) in vivo testing, and...

  49. Cloning

    Type
    Guide
    ...add restriction sites to any piece of DNA during PCR amplification, allowing for it to then be digested...a single adenosine (A) overhang on the 3' end of PCR products. The complementary thymine (T) comes from...an incredibly quick and easy way to clone a fresh PCR product into a plasmid. The major disadvantage of...are also available, allowing for rapid cloning of PCR products into donor plasmids without the need for...used, so if properly designed, any insert fragment (PCR product or synthesized oligo) with appropriate overhangs...add). Complementary overhangs are built into the PCR primers for the insert, based on the destination ...
Showing: 101 - 150 of 207 results