From 429eb186f237c00de7fe2ea191bf73f2bc06c372 Mon Sep 17 00:00:00 2001 From: Charles Plessy Date: Thu, 19 Apr 2018 10:51:18 +0900 Subject: [PATCH] Reorganise and add pseudo-random primers. --- tags/reverse_transcription.mdwn | 25 +++++++++++++++---------- 1 file changed, 15 insertions(+), 10 deletions(-) diff --git a/tags/reverse_transcription.mdwn b/tags/reverse_transcription.mdwn index f6230ba7..fcc03633 100644 --- a/tags/reverse_transcription.mdwn +++ b/tags/reverse_transcription.mdwn @@ -1,13 +1,17 @@ [[!meta title="pages tagged reverse transcription"]] -(redaction in progress) +The reverse transcriptase +========================= -Additives that increase reaction performance: +_(redaction in progress)_ + +## Additives that increase reaction performance: - Actinomycin D ([[Perocchi et al., 2007|biblio/17897965]]). - - T4 bacteriophage gene 32 protein (T4gp32, [[Kenzelmann _et al._, 2004|biblio/15028277]], [[Piché _et al._, 2005|biblio/16461948]]). + - [[T4 bacteriophage gene 32 protein|ssbp]] (T4gp32, [[Kenzelmann _et al._, 2004|biblio/15028277]], + [[Piché _et al._, 2005|biblio/16461948]]). -Reverse-transcriptases have a DNA-dependent DNA polymerase activity. +## DNA-dependent DNA polymerase activity. - It is utilised in [[template_switching]] methods to add linkers to first-strand cDNAs. - It is also a source of antisense artefacts when the RT makes a second-strand cDNA @@ -15,16 +19,17 @@ Reverse-transcriptases have a DNA-dependent DNA polymerase activity. but not RNA-dependent polymerase activity and is used to suppress these artefacts ([[Perocchi et al., 2007|biblio/17897965]], [[Kanamori-Katayama et al., 2011|biblio/21596820]]). -Reverse-transcriptases tolerate terminal mismatches: [[Mizuno et al., 1999|biblio/9973624]]. +## Reverse-transcriptases tolerate terminal mismatches: + + - Reported by [[Mizuno et al., 1999|biblio/9973624]]. + - Utilised in [[Arnaud et al., 2015|biblio/27071605]] to reduce priming or + ribosomal or hemoglobin RNA. -Reverse-transcription primers: +## Reverse-transcription primers: - "N15" random pentadecamers: [[Stangegaard et al., 2006|biblio/16708763]]. - Multi-targeted primers (MTP): [[Adomas et al., 2010|biblio/20716356]]. - "Not-so random" (NSR) primers: [[Armour et al., 2009|biblio/19668204]]. - - ... and many more (pseudo-random, ...) - -[[Single-strand binding proteins|ssbp]] are sometimes added to improve reverse -transcriptions. + - "Pseudo-random" primers: [[Arnaud et al., 2015|biblio/27071605]]. [[!inline pages="tagged(reverse_transcription)" actions="no" limit=0]] -- 2.47.3