ddPCR experiments were performed with a QX200 ddPCR system (Bio-Rad) and ddPCR Eva Green Super Mix kit (Bio-Rad), according to the manufacturer’s protocol. Briefly, cDNA (1.5 μl per sample) and the corresponding primer pairs (0.165 μM; Sigma) were added to the Eva Green Super Mix at a final volume of 20 μl. Then, droplets were generated with the aid of a QX200 droplet generator by adding 70 μl of the ddPCR oil. The emulsion product (40 μl) was subsequently transferred to a 96-well plate for PCR reaction. Finally, the samples were read using a Bio-Rad QX200 droplet reader. Data were analyzed with a QuantaSoft software and expressed as number of copies per micrograms of RNA [27 (link)]. Forward (F) and reverse (R) primers were designed for each gene by using the Beacon Designer software (Premier Biosoft). Primer sequences used are shown in Table 1 .
Ddpcr evagreen supermix kit
Manufactured by Bio-Rad
The DdPCR EvaGreen Supermix kit is a reagent designed for use with droplet digital PCR (ddPCR) systems. It contains all the necessary components, including the EvaGreen dye, to perform digital PCR amplification and quantification of DNA or RNA targets.
Automatically generated - may contain errors
Lab products found in correlation
Qx200 droplet reader, by Bio-Rad (4 mentions)
Quantasoft software, by Bio-Rad (3 mentions)
Iscript advanced cdna synthesis kit, by Bio-Rad (3 mentions)
Kapa duel indexed adapters, by Roche (2 mentions)
High sensitivity dna screentape, by Agilent Technologies (2 mentions)
Kapa rna hyperprep kit with riboerase hmr, by Roche (2 mentions)
Hiseq 4000 platform, by Illumina (2 mentions)
Tapestation 4200, by Agilent Technologies (2 mentions)
Primer pair, by Merck Group (1 mentions)
Qx200 ddpcr system, by Bio-Rad (1 mentions)
Kapa library quantification kit, by Roche (1 mentions)
4 protocols using ddpcr evagreen supermix kit
1
Droplet Digital PCR Quantification of Gene Expression
2
Reverse Transcription and ddPCR Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
cDNA was reverse transcribed from 10 ng total RNA using an iScript Advanced cDNA synthesis kit (Bio-Rad). ddPCR was performed by using ddPCR EvaGreen Supermix kit (Bio-Rad) on a QX200 Droplet Reader (Bio-Rad) based on the manufacturer’s protocol. Results were analyzed using QuantaSoft software (Bio-Rad).
3
Transcriptomic Profiling of Sorted Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For RNA-seq of samples from sorted neurons (the sort group) and whole worms (the whole group), libraries were prepared using KAPA RNA HyperPrep kit with RiboErase (HMR) (KAPA biosystems) according to the manufacturer's protocol. The RNA input was 150 ng and fragmentation conditions were 85 o C for 5 min. Barcodes were introduced to each sample using KAPA duel-indexed adapters (KAPA biosystems). Length distribution of each library was determined by TapeStation 4200 (Agilent) using High Sensitivity DNA ScreenTape (Agilent). Libraries were quanti ed by KAPA library quanti cation kit (KAPA biosystems) and then multiplexed and sequenced on Illumina Hiseq 4000 platform to obtain 150 nt paired-end reads.
Droplet digital PCR (ddPCR) cDNA was reverse transcribed from 10 ng total RNA using an iScript Advanced cDNA synthesis kit (Bio-Rad). ddPCR was performed by using ddPCR EvaGreen Supermix kit (Bio-Rad) on a QX200 Droplet Reader (Bio-Rad) based on the manufacturer's protocol. Results were analyzed using QuantaSoft software (Bio-Rad).
Droplet digital PCR (ddPCR) cDNA was reverse transcribed from 10 ng total RNA using an iScript Advanced cDNA synthesis kit (Bio-Rad). ddPCR was performed by using ddPCR EvaGreen Supermix kit (Bio-Rad) on a QX200 Droplet Reader (Bio-Rad) based on the manufacturer's protocol. Results were analyzed using QuantaSoft software (Bio-Rad).
4
RNA-seq and ddPCR for Sorted Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For RNA-seq of samples from sorted neurons (the sort group) and whole worms (the whole group), libraries were prepared using KAPA RNA HyperPrep kit with RiboErase (HMR) (KAPA biosystems) according to the manufacturer's protocol. The RNA input was 150 ng and fragmentation conditions were 85 o C for 5 min. Barcodes were introduced to each sample using KAPA duel-indexed adapters (KAPA biosystems). Length distribution of each library was determined by TapeStation 4200 (Agilent) using High Sensitivity DNA ScreenTape (Agilent). Libraries were quantified by KAPA library quantification kit (KAPA biosystems) and then multiplexed and sequenced on Illumina Hiseq 4000 platform to obtain 150 nt paired-end reads.
Droplet digital PCR (ddPCR) cDNA was reverse transcribed from 10 ng total RNA using an iScript Advanced cDNA synthesis kit (Bio-Rad). ddPCR was performed by using ddPCR EvaGreen Supermix kit (Bio-Rad) on a QX200 Droplet Reader (Bio-Rad) based on the manufacturer's protocol. Results were analyzed using QuantaSoft software (Bio-Rad).
Droplet digital PCR (ddPCR) cDNA was reverse transcribed from 10 ng total RNA using an iScript Advanced cDNA synthesis kit (Bio-Rad). ddPCR was performed by using ddPCR EvaGreen Supermix kit (Bio-Rad) on a QX200 Droplet Reader (Bio-Rad) based on the manufacturer's protocol. Results were analyzed using QuantaSoft software (Bio-Rad).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!