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Iqbal,Saima; Bashir,Aftab; Naseer,Hafiza Masooma; Ahmed,Moddassir; Malik,Kauser A. |
Cotton fibers are differentiated, non-dividing cells that originate from the epidermal layer of developing ovules. To identify genes involved in cotton fiber development, we performed non-radioactive differential display reverse transcriptase PCR (DDRT-PCR) on the purified mRNA. This technique was tested on mRNA isolated from five different developmental stages of cotton fiber including 0, 5, 10, 15 and 20 DPA (days after pollination). The mRNA purified from total RNA was reversibly transcribed using three anchored oligo-dT primers. Polymerase chain reaction (PCR) amplification of each cDNA preparation was carried out in combination with seven arbitrary primers. The amplified products were resolved on 1% agarose gel containing ethidium bromide. DNA was... |
Tipo: Journal article |
Palavras-chave: Cotton fiber; Differentially expressed transcripts; Gossypium hirsutum; Non-radioactive. |
Ano: 2008 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582008000100005 |
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LeBlanc-Straceski,Janine; Sobrado,Pablo; Betz,Sharon; Zerfas,Julie; Morgan,Karen. |
A PCR based strategy was developed to screen a Xenopus oocyte λgt10 cDNA library. The PCR-based lift pool (LP) method follows the same two tiered strategy as conventional screening of phage libraries by filter hybridization. Two rounds of plating, one at high density to detect the clone, and one at low density to purify the clone to homogeneity, are performed. In the first round, lysates from high density plates, termed plate pools (PP), serve as template for PCR. In the second round, phage particles adhering to plaque lifts of low density plates are washed off nitrocellulose filters to create LPs, which are used as template for PCR. The integrity of the plaques on the low-density plates is preserved. Once a positive LP has been identified,... |
Tipo: Journal article |
Palavras-chave: CDNA; Lambda; Non-radioactive; PCR; Plaque; Screening. |
Ano: 2006 |
URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582006000400012 |
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