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

...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...

...Development and optimization of a real-time quantitative PCR-based method for the titration of AAV-2 vector stocks...

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

... a blood draw, tail clip, or toe clip) and using PCR with the appropriate primers to detect your targeting...

...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 ...

...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 ...

...screen for this specific recombination outcome using PCR and then generate a monoclonal cell line with the...

... have DNA left over for other applications, like PCR, cloning, transfection, or long-term storage. But...

...insertions/deletions using a mismatch repair assay? Or is PCR followed by gel electrophoresis and/or next-generation...

...the purified virus should be ~1.3 Quantitative PCR: the number of viral DNA packaged in virions is determined...

...efficacy, we have used both Western blotting and PCR, depending on the level of endogenous expression....

...it’s so easy once you are in the lab. There is no PCR, no gel purification; the whole cloning process is...

...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...

...titrated following purification using qPCR or digital droplet PCR. Insect cells Insect cells are a less...

...both edited and unedited (control) populations is PCR-amplified and Sanger sequenced. ICE compares these...

...bound genomic locus, which they then confirmed by PCR. Subsequent mass spectroscopy studies allowed for...

...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...

... selection. Screening of transformants by colony PCR confirmed the insertion of the Lambda insert at the...

...plate overnight Screen colonies phenotypically or by PCR/sequencing While CRISPR-transposons are powerful...