Real-time PCR was performed as described above using Kapa HiFi HotStart ReadyMix as directed by the manufacturer (Supplementary Table 7 : 5.8kb_forward/reverse, 5.8kb_f_alt/r_alt), depending on the desired target, and the following thermal cycling protocol: 95°C for 180 sec, repeat [98°C for 20 sec, 65°C for 15 sec, 72°C for 180 sec]. Samples were loaded on a 1% agarose gel and run for 1 hour at 100 V. Bands of the desired size were size selected and purified (Supplementary Fig. 9 ). A second, four-fold larger volume PCR using the same conditions was then run using the size-selected sample. The PCR products were pooled for each individual and purified using Agencourt AMPure magnetic beads, following the manufacturer’s protocol. Samples were eluted in 50 μl and loaded directly into the Covaris Adaptive Focused Acoustics machine. Standard shotgun libraries were then prepared for paired-end (2 × 250 bp) Illumina MiSeq sequencing (sequencing primers in Supplementary Table 7 : MiSeq_p7 and MiSeq_p5).
Adaptive focused acoustics machine
Manufactured by Covaris
The Adaptive Focused Acoustics machine is a laboratory equipment designed for sample preparation and processing. It utilizes focused acoustic energy to disrupt and homogenize samples, such as cells, tissues, or other materials, without the use of physical forces or mechanical disruption. The machine is capable of precise and controlled sample processing, ensuring consistent and reproducible results.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using adaptive focused acoustics machine
1
Amplification and Sequencing of Large Genomic Fragments
2
Amplification and Sequencing of Large Genomic Fragments
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Real-time PCR was performed as described above using Kapa HiFi HotStart ReadyMix as directed by the manufacturer (Supplementary Table 7 : 5.8kb_forward/reverse, 5.8kb_f_alt/r_alt), depending on the desired target, and the following thermal cycling protocol: 95°C for 180 sec, repeat [98°C for 20 sec, 65°C for 15 sec, 72°C for 180 sec]. Samples were loaded on a 1% agarose gel and run for 1 hour at 100 V. Bands of the desired size were size selected and purified (Supplementary Fig. 9 ). A second, four-fold larger volume PCR using the same conditions was then run using the size-selected sample. The PCR products were pooled for each individual and purified using Agencourt AMPure magnetic beads, following the manufacturer’s protocol. Samples were eluted in 50 μl and loaded directly into the Covaris Adaptive Focused Acoustics machine. Standard shotgun libraries were then prepared for paired-end (2 × 250 bp) Illumina MiSeq sequencing (sequencing primers in Supplementary Table 7 : MiSeq_p7 and MiSeq_p5).
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!