3 5 klenow fragment

Manufactured by New England Biolabs

The 3′−5′ Klenow Fragment is a DNA polymerase enzyme derived from the Klenow fragment of DNA Polymerase I. It possesses 5′→3′ polymerase activity and 3′→5′ exonuclease activity, but lacks the 5′→3′ exonuclease activity of the full-length DNA Polymerase I.

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Lab products found in correlation

Poli klenow fragment, by New England Biolabs (2 mentions) Nebnext ultraii q5 kit, by New England Biolabs (2 mentions) T4 polynucleotide kinase, by New England Biolabs (2 mentions) T4 dna polymerase, by New England Biolabs (2 mentions) Sonicator, by Covaris (2 mentions) T4 dna ligase, by New England Biolabs (2 mentions) Quick ligase, by New England Biolabs (2 mentions)

Close spelling variants of this product

Klenow Fragment (3’ to 5’ Exo-), Klenow Fragment (3′ -> 5′ exo-) Klenow Fragment 3′-5′ exo-nuclease Klenow fragment (3′–5′ exo-) polymerase (KF polymerase) Klenow Fragment (3′→5′ exo-minus, Klenow fragment

2 protocols using 3 5 klenow fragment

Whole Genome Sequencing Library Preparation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

20 0- to 3-day-old w1118 males or females were collected on dry ice and then homogenized using an electric pestle. Qia-Amp DNA Micro kit was used according to instructions. DNA was diluted to 40 ng/µl in 55 µl of Elution Buffer and sheared in a Covaris sonicator with settings as follows: 10% duty cycle, 2.0 intensity, 200 cycles per burst, 1 cycle, and 45 s process time.
WGS library generation protocol was adapted from the Marshall-Lab DamID-seq protocol^{78 (link)} available at marshall-lab.org. Briefly, sheared DNA was purified with homemade purification beads. End repair was performed with T4 DNA Ligase (NEB M0202S), T4 DNA Polymerase (NEB M0203S), PolI Klenow fragment (NEB M0210S), and T4 Polynucleotide kinase (NEB M0201S). Adenylation was performed with 3′−5′ Klenow Fragment (NEB M0212L). Adaptors were ligated with NEB Quick Ligase for 10 min at 30 °C before two rounds of cleanup with homemade beads. NEBNext UltraII Q5 kit (NEB M0544) was used for PCR enrichment. A final round of cleanup with homemade beads was performed before quantification and sequencing.

Lawlor M.A., Cao W, & Ellison C.E. (2021). A transposon expression burst accompanies the activation of Y-chromosome fertility genes during Drosophila spermatogenesis. Nature Communications, 12, 6854.

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Whole Genome Sequencing Library Preparation

Check if the same lab product or an alternative is used in the 5 most similar protocols

20 0-to 3-day old w1118 males or females were collected on dry ice and then homogenized using an electric pestle. Qia-Amp DNA Micro kit was used according to instructions. DNA was diluted to 40 ng/ul in 55 ul of Elution Buffer and sheared in a Covaris sonicator with settings as f ollows: 10% duty cycle, 2.0 intensity, 200 cycles per burst, 1 cycle, 45 second process time.
WGS library generation protocol was adapted from the Marshall Lab DamID-seq protocol available at marshalllab.org (Marshall et al. 2016) . Briefly, sheared DNA was purified with homemade purification beads. End repair was performed with T4 DNA Ligase (NEB M0202S), T4 DNA Polymerase (NEB M0203S) , PolI Klenow fragment (NEB M0210S), T4 Polynucleotide kinase (NEB M0201S). Adenylation was performed with 3'-5' Klenow Fragment (NEB M0212L). Adaptors were ligated with NEB Quick Ligase for 10 minutes at 30℃ before two rounds of cleanup with homemade beads. NEBNext UltraII Q5 kit (NEB M0544) was used for PCR enrichment. A final round of cleanup with homemade beads was performed before quantification and sequencing.

Lawlor M.A., Cao W., & Ellison C. (2021). A burst of transposon expression accompanies the activation of Y chromosome fertility genes during Drosophila spermatogenesis.

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