The frozen samples were lysed to release crude cell protein on ice in TRAPeze® 1 × CHAPS lysis buffer (Millipore, Boston, USA). After quantitation, 1 μg of protein lysate was used for each telomerase activity assay using TRAPeze® RT Telomerase Detection Kit (Millipore, Boston, USA). Briefly, the PCR was conducted in a volume of 12.5 μl including 2.5 μl of 5 × TRAPeze® RT Reaction Mix, 8.8 μl of nuclease-free water, 0.2 μl of Taq polymerase, and 1 μl of the protein sample. A dilution series of TSR8 control template was to serve as a standard curve. Reactions were performed in triplicate. The thermal cycling conditions comprised one cycle for 30 min at 30°C, 2 min at 95°C, followed by 45 cycles of 15 s at 94°C, 1 min at 59°C, and 10 s at 45°C. According to the protocol, the positive telomerase activity should be above background at a dilution of 0.04 amols (Log copy number = 4.38) of TSR8 control template.
Trapeze 1 chaps lysis buffer
Manufactured by Merck Group
Sourced in United States
TRAPeze® 1 × CHAPS lysis buffer is a solution used for cell lysis and protein extraction. It contains the zwitterionic detergent CHAPS, which aids in the solubilization of proteins from biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Gelred, by Biotium (2 mentions)
Trapeze rt telomerase detection kit, by Merck Group (1 mentions)
Dr gentle precipitation carrier, by Takara Bio (1 mentions)
Uracil dna glycosylase, by New England Biolabs (1 mentions)
Nebuffer 2, by New England Biolabs (1 mentions)
Sybr gold, by Thermo Fisher Scientific (1 mentions)
T4 dna ligase, by New England Biolabs (1 mentions)
Trapeze telomerase detection kit, by Merck Group (1 mentions)
Bradford assay, by Bio-Rad (1 mentions)
3 protocols using trapeze 1 chaps lysis buffer
1
Quantifying Telomerase Activity in Cells
2
DNA Repair Enzyme-Based Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Uracil-DNA
glycosylase (UDG), human alkyladenine glycosylase (hAAG), uracil glycosylase
inhibitor (UGI), T4 DNA ligase, exonuclease I (Exo I), exonuclease
III (Exo III), endonuclease IV (Endo IV), and 10× NEBuffer 2
(500 mM NaCl, 100 mM Tris-HCl, 100 mM MgCl2, 10 mM dithiothreitol)
were purchased from New England Biolabs (Ipswich, MA, USA). Dr. GenTLE
Precipitation Carrier was purchased from Takara Biotechnology Co.,
Ltd. (Dalian, China). GelRed was purchased from Biotium (Shanghai,
China). 10× SYBR Green I was purchased from Zeesan (Xiamen, China).
SYBR Gold was purchased from Invitrogen (CA, USA). NxGen phi29 DNA
polymerase was purchased from Lucigen (WI, USA). TRAPeze 1× CHAPS
lysis buffer was purchased from Millipore Corp. (Billerica, MA, USA).
All other chemicals were analytical grade and used as received. All
ultrapure water (18.2 MΩ) used in this work was acquired through
a Milli-Q water purification system (Billerica, MA). All oligonucleotides
used in the assay inTable 1 were synthesized by Takara Biotechnology Co., Ltd. (Dalian,
China).
glycosylase (UDG), human alkyladenine glycosylase (hAAG), uracil glycosylase
inhibitor (UGI), T4 DNA ligase, exonuclease I (Exo I), exonuclease
III (Exo III), endonuclease IV (Endo IV), and 10× NEBuffer 2
(500 mM NaCl, 100 mM Tris-HCl, 100 mM MgCl2, 10 mM dithiothreitol)
were purchased from New England Biolabs (Ipswich, MA, USA). Dr. GenTLE
Precipitation Carrier was purchased from Takara Biotechnology Co.,
Ltd. (Dalian, China). GelRed was purchased from Biotium (Shanghai,
China). 10× SYBR Green I was purchased from Zeesan (Xiamen, China).
SYBR Gold was purchased from Invitrogen (CA, USA). NxGen phi29 DNA
polymerase was purchased from Lucigen (WI, USA). TRAPeze 1× CHAPS
lysis buffer was purchased from Millipore Corp. (Billerica, MA, USA).
All other chemicals were analytical grade and used as received. All
ultrapure water (18.2 MΩ) used in this work was acquired through
a Milli-Q water purification system (Billerica, MA). All oligonucleotides
used in the assay in
China).
3
Quantitative Telomerase Activity Assay
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!